subject:(recombinant protein expression AND purification)

University of Sydney

1. Wu, Jiadai. Haliotis rubra hemocyanin: sequencing, recombinant expression and investigation of the binding activities of expressed function units with HSV-1 .

Degree: 2017, University of Sydney

URL: http://hdl.handle.net/2123/17792

► Hemocyanin (HrH) has been shown to possess antiviral activity against HSV-1 by selectively binding to viral glycoproteins (gB, gC, gD). Its large size however precluded… (more)

Subjects/Keywords: Haliotis rubra hemocyanin; Herpes Simplex Virus type 1; Gene Clone; Protein recombinant expression; Protein purification; Protein-protein interaction

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Wu, J. (2017). Haliotis rubra hemocyanin: sequencing, recombinant expression and investigation of the binding activities of expressed function units with HSV-1 . (Thesis). University of Sydney. Retrieved from http://hdl.handle.net/2123/17792

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Wu, Jiadai. “Haliotis rubra hemocyanin: sequencing, recombinant expression and investigation of the binding activities of expressed function units with HSV-1 .” 2017. Thesis, University of Sydney. Accessed February 28, 2021. http://hdl.handle.net/2123/17792.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Wu, Jiadai. “Haliotis rubra hemocyanin: sequencing, recombinant expression and investigation of the binding activities of expressed function units with HSV-1 .” 2017. Web. 28 Feb 2021.

Vancouver:

Wu J. Haliotis rubra hemocyanin: sequencing, recombinant expression and investigation of the binding activities of expressed function units with HSV-1 . [Internet] [Thesis]. University of Sydney; 2017. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/2123/17792.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Wu J. Haliotis rubra hemocyanin: sequencing, recombinant expression and investigation of the binding activities of expressed function units with HSV-1 . [Thesis]. University of Sydney; 2017. Available from: http://hdl.handle.net/2123/17792

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Michigan Technological University

2. Hopson, Sarah. Heterologous Expression and Purification of Full-Length Human Polybromo-1 Protein.

Degree: PhD, Department of Chemistry, 2017, Michigan Technological University

URL: https://digitalcommons.mtu.edu/etdr/436

► Over the past decade, it has become apparent that the human polybromo-1 protein (BAF180) has a critical role in cancer. BAF180 is known to… (more)

▼ Over the past decade, it has become apparent that the human polybromo-1 protein (BAF180) has a critical role in cancer. BAF180 is known to be a driver mutation in clear cell renal cell carcinoma, where it has been found to be mutated in approximately 40% of cases. Mutations have also been found in several other cancers, including intrahepatic cholangiocarcinomas and epithelioid sarcomas. BAF180 is the chromatin targeting subunit of the PBAF (Polybromo-associated BRG1-associated factor) chromatin remodeling complex, a role facilitated by its nine domains: six bromodomains, which recognize and bind to acetylated lysines on histones; two BAH (bromo-adjacent homology) domains, found to be critical for PCNA ubiquitination following DNA damage; and one HMG (high mobility group) box, the DNA binding component. Furthermore, proper expression of BAF180 has also been linked to cardiac development and cell cycle regulation. Despite these associations, the molecular level interactions of full-length BAF180 have yet to be studied; only the phenomenological effects of BAF180 deficiency/mutation have been studied. It is crucial that we understand the binding dynamics and specificities of wild type and mutated BAF180, since it is the recognition component of PBAF. Expression of the recombinant full-length BAF180 protein has been difficult because of the complex nature of this protein, its unusual codon usage, and size. After E. coli expression failed, other expression systems were investigated and the yeast Pichia pastoris was chosen. Pichia was chosen for several reasons: its codon usage is similar to that of BAF180 and it is a eukaryotic system possessing eukaryotic protein folding machinery and capable of performing post-translational modifications. Under control of the GAP promoter, full-length BAF180 has been successfully expressed in Pichia pastoris. This is the first time that full-length BAF180 has been cloned and expressed in a heterologous host. It was purified using anion exchange chromatography. The ability to express and purify full-length BAF180 is a huge first step towards increasing the understanding of the molecular mechanisms of this protein and its association with cancer development. Advisors/Committee Members: Martin Thompson.

Subjects/Keywords: recombinant protein expression and purification; human polybromo-1; BAF180; PBRM1; hPB1; Pichia pastoris; Biochemistry; Molecular Biology

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Hopson, S. (2017). Heterologous Expression and Purification of Full-Length Human Polybromo-1 Protein. (Doctoral Dissertation). Michigan Technological University. Retrieved from https://digitalcommons.mtu.edu/etdr/436

Chicago Manual of Style (16^th Edition):

Hopson, Sarah. “Heterologous Expression and Purification of Full-Length Human Polybromo-1 Protein.” 2017. Doctoral Dissertation, Michigan Technological University. Accessed February 28, 2021. https://digitalcommons.mtu.edu/etdr/436.

MLA Handbook (7^th Edition):

Hopson, Sarah. “Heterologous Expression and Purification of Full-Length Human Polybromo-1 Protein.” 2017. Web. 28 Feb 2021.

Vancouver:

Hopson S. Heterologous Expression and Purification of Full-Length Human Polybromo-1 Protein. [Internet] [Doctoral dissertation]. Michigan Technological University; 2017. [cited 2021 Feb 28]. Available from: https://digitalcommons.mtu.edu/etdr/436.

Council of Science Editors:

Hopson S. Heterologous Expression and Purification of Full-Length Human Polybromo-1 Protein. [Doctoral Dissertation]. Michigan Technological University; 2017. Available from: https://digitalcommons.mtu.edu/etdr/436

3. Belchior, Gustavo Gross. Geração de clones de células HEK293 superprodutores de isoformas recombinantes de VEGF-A (Fator de Crescimento Endotelial Vascular A) humano visando à produção de biofármacos para terapia molecular e engenharia tecidual.

Degree: PhD, Bioquímica, 2014, University of São Paulo

URL: http://www.teses.usp.br/teses/disponiveis/46/46131/tde-01102014-145602/ ;

► Os primeiros vasos sanguíneos do embrião de vertebrados, formados de novo a partir de células originadas da mesoderme, originam os vasos linfáticos em um processo… (more)

▼ Os primeiros vasos sanguíneos do embrião de vertebrados, formados de novo a partir de células originadas da mesoderme, originam os vasos linfáticos em um processo denominado vasculogênese. Já no adulto, novos vasos são formados principalmente através da angiogênese (ou linfoangiogênese) a partir da vasculatura pré-existente. Em indivíduos saudáveis, a arquitetura vascular é relativamente estática, sendo que o excesso ou a insuficiência de vasos são comumente relacionados à angiogênese patológica, vinculada a diversas doenças, como câncer, degeneração macular relacionada à idade, isquemia de membros e muitas outras. Dessa forma, o controle local da densidade de vasos sanguíneos torna-se interessante para o tratamento de condições patológicas visando melhoria de prognóstico e cura. Dentre os diversos fatores de crescimento conhecidos, o fator de crescimento endotelial vascular, VEGF, destaca-se como o principal regulador do processo de angiogênese através de isoformas pró-angiogênicas (VEGFxxx) e antiangiogênicas (VEGFxxxb) do gene VEGF-A. Consequentemente, as proteínas codificadas por esse gene constituem um alvo com alto potencial terapêutico. No presente estudo, propusemos a produção das isoformas proteicas recombinantes rhVEGF165, rhVEGF165b e rhVEGF121, oriundas do gene VEGF-A humano, visando à geração de biofármacos que podem ser utilizados para terapia molecular e engenharia tecidual. As sequências codificadoras das isoformas rhVEGF165 e rhVEGF121 foram amplificadas a partir do cDNA total sintetizado de amostras de RNA total de pulmão humano, enquanto que a da isoforma rhVEGF165b foi gerada através da mutação sítio-dirigida da sequência de rhVEGF165. As sequências foram clonadas no vetor de clonagem pGEM®-T Easy, sendo em seguida subclonadas no vetor pLV-eGFP, um vetor plasmidial de transferência lentiviral, que permite a expressão de transgenes em células de mamíferos e, também, da proteína repórter eGFP. Células humanas HEK293 em cultura aderente foram independentemente cotransfectadas com cada uma das construções geradas (pLV-rhVEGF165, pLV-rhVEGF165b e pLV-rhVEGFl21) juntamente com o vetor pTK-Hyg na proporção de 40:1 (m/m), viabilizando a seleção dos transfectantes com o antibiótico higromicina B, além da detecção de eGFP. Os clones celulares superprodutores das proteínas de interesse foram avaliados quanto à cinética de expressão em meio carente de soro e adaptados para a cultura em suspensão estática na presença de meio na ausência de componentes derivados de animais, demostrando a capacidade de expressão das isoformas rhVEGFs nestas condições de cultivo. Para o nosso conhecimento, este é o primeiro trabalho a descrever a expressão de isoformas de VEGF-A em células HEK293 mantidas em suspensão. As isoformas rhVEGF165 e rhVEGF165b foram purificadas por cromatografia de afinidade a heparina, a partir do meio condicionado pelos clones superprodutores gerados. Ensaios in vitro, utilizando o AngioPhaseTM Kit, e in vivo, através do ensaio da membrana corioalantoide em embriões de galinha (CAM Assay), ambos… Advisors/Committee Members: Sogayar, Mari Cleide.

Subjects/Keywords: Angiogênese; Angiogenesis; Células HEK293; HEK293 cells; Heterologous expression system; Protein purification; Proteínas recombinantes; Purificação de proteínas; Recombinant proteins; Sistema de expressão heterólogo; VEGF; VEGF

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Belchior, G. G. (2014). Geração de clones de células HEK293 superprodutores de isoformas recombinantes de VEGF-A (Fator de Crescimento Endotelial Vascular A) humano visando à produção de biofármacos para terapia molecular e engenharia tecidual. (Doctoral Dissertation). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/46/46131/tde-01102014-145602/ ;

Chicago Manual of Style (16^th Edition):

Belchior, Gustavo Gross. “Geração de clones de células HEK293 superprodutores de isoformas recombinantes de VEGF-A (Fator de Crescimento Endotelial Vascular A) humano visando à produção de biofármacos para terapia molecular e engenharia tecidual.” 2014. Doctoral Dissertation, University of São Paulo. Accessed February 28, 2021. http://www.teses.usp.br/teses/disponiveis/46/46131/tde-01102014-145602/ ;.

MLA Handbook (7^th Edition):

Belchior, Gustavo Gross. “Geração de clones de células HEK293 superprodutores de isoformas recombinantes de VEGF-A (Fator de Crescimento Endotelial Vascular A) humano visando à produção de biofármacos para terapia molecular e engenharia tecidual.” 2014. Web. 28 Feb 2021.

Vancouver:

Belchior GG. Geração de clones de células HEK293 superprodutores de isoformas recombinantes de VEGF-A (Fator de Crescimento Endotelial Vascular A) humano visando à produção de biofármacos para terapia molecular e engenharia tecidual. [Internet] [Doctoral dissertation]. University of São Paulo; 2014. [cited 2021 Feb 28]. Available from: http://www.teses.usp.br/teses/disponiveis/46/46131/tde-01102014-145602/ ;.

Council of Science Editors:

Belchior GG. Geração de clones de células HEK293 superprodutores de isoformas recombinantes de VEGF-A (Fator de Crescimento Endotelial Vascular A) humano visando à produção de biofármacos para terapia molecular e engenharia tecidual. [Doctoral Dissertation]. University of São Paulo; 2014. Available from: http://www.teses.usp.br/teses/disponiveis/46/46131/tde-01102014-145602/ ;

University of Victoria

4. Jantzen, Roni Rebecca. Expression of human α-N-Acetylglucosaminidase in Sf9 insect cells: effect of cryptic splice site removal and native secretion-signaling peptide addition.

Degree: Dept. of Biology, 2011, University of Victoria

URL: http://hdl.handle.net/1828/3455

► Human α-N-Acetylglucosaminidase (Naglu) is a lysosomal acid hydrolase implicated in tthe rare metabolic storage disorder known as mucopolysaccharidosis type IIIB (MPS IIIB; also Sanfilippo syndrome… (more)

Subjects/Keywords: α-N-Acetylglucosaminidase; Sf9; insect cells; secretion signal peptide; lysosomal enzyme; mucopolysaccharidosis; MPS IIIB; sanfilippo syndrome; protein expression; protein transduction domain; splice site; cryptic splicing; site-directed mutagenesis; Naglu; cDNA; protein purification; hydrophobic interaction chromatography; FPLC; recombinant human protein; fusion protein; Tat-PTD

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APA (6^th Edition):

Jantzen, R. R. (2011). Expression of human α-N-Acetylglucosaminidase in Sf9 insect cells: effect of cryptic splice site removal and native secretion-signaling peptide addition. (Masters Thesis). University of Victoria. Retrieved from http://hdl.handle.net/1828/3455

Chicago Manual of Style (16^th Edition):

Jantzen, Roni Rebecca. “Expression of human α-N-Acetylglucosaminidase in Sf9 insect cells: effect of cryptic splice site removal and native secretion-signaling peptide addition.” 2011. Masters Thesis, University of Victoria. Accessed February 28, 2021. http://hdl.handle.net/1828/3455.

MLA Handbook (7^th Edition):

Jantzen, Roni Rebecca. “Expression of human α-N-Acetylglucosaminidase in Sf9 insect cells: effect of cryptic splice site removal and native secretion-signaling peptide addition.” 2011. Web. 28 Feb 2021.

Vancouver:

Jantzen RR. Expression of human α-N-Acetylglucosaminidase in Sf9 insect cells: effect of cryptic splice site removal and native secretion-signaling peptide addition. [Internet] [Masters thesis]. University of Victoria; 2011. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/1828/3455.

Council of Science Editors:

Jantzen RR. Expression of human α-N-Acetylglucosaminidase in Sf9 insect cells: effect of cryptic splice site removal and native secretion-signaling peptide addition. [Masters Thesis]. University of Victoria; 2011. Available from: http://hdl.handle.net/1828/3455

Université de Grenoble

5. Ganne, Géraldine. Etudes structurales et fonctionnelles de lectines et d'adhésines chez Pseudomonas aeruginosa : Structural and functional characterisation of lectines and adhesins from Pseudomonas aeruginosa.

Degree: Docteur es, Sciences de la vie, 2013, Université de Grenoble

URL: http://www.theses.fr/2013GRENV005

►

Pseudomonas aeruginosa est un pathogène opportuniste responsable de nombreuses maladies nosocomiales chez les patients immunodéprimés ainsi que d'infections graves chez les patients atteints de la… (more)

▼

Pseudomonas aeruginosa est un pathogène opportuniste responsable de nombreuses maladies nosocomiales chez les patients immunodéprimés ainsi que d'infections graves chez les patients atteints de la mucoviscidose (CF). La colonisation des voies respiratoires des patients CF est souvent mortelle car une fois installée, cette bactérie est difficile à éradiquer et provoque le déclin des fonctions respiratoires des patients. L'antibiothérapie devient inefficace face au développement de souches multi-résistantes et sa capacité à former un biofilm. L'étape cruciale initiant l'infection ou la formation du biofilm est l'adhésion durant laquelle des interactions spécifiques lectines/oligosaccharides permettent la fixation de la bactérie à la surface de la cellule hôte. Bloquer l'adhésion serait un moyen de lutter contre l'infection. Dans l'approche d'une thérapie anti-adhésive, plusieurs lectines impliquées dans l'adhésion et l'élaboration du biofilm sont prises comme cibles. Dans un premier temps, des lectines fimbriales putatives identifiées récemment, CupB6 et CupE6, ont fait l'objet d'une étude. Des essais d'expression, de purification et de cristallisation ont été réalisés dans l'objectif de résoudre leur structure cristallographique. Une étude visant à identifier le ligand naturel de CupB6 a également été entreprise. Puis une étude a été menée pour caractériser le potentiel d'inhibition de plusieurs molécules dérivées du galactose sur la lectine soluble, PA-IL. Certaines de ces molécules pourraient être utilisées comme glycomimétiques offrant une alternative aux antibiotiques. Une étude par microcalorimétrie et cristallographie aux rayons X a permis d'étudier la spécificité d'une lectine de légumineuse, PELa.

P. aeruginosa is an opportunistic pathogenic bacterium responsible for numerous nosocomial infections in immunosuppressed patients. It is the first mortal pathogen in cystic fibrosis (CF) patients. The invasion of the respiratory tract of CF patients by the bacterium is often lethal because it is hard to eradicate and it rapidly impairs the respiratory functions of the patients. None of the current antibiotherapy procedures are efficient against multiresistant, biofilm forming P. aeruginosa. The first step leading to infection or biofilm formation involves the initial adhesion of bacterial cells to the host pulmonary cells via specific lectin/oligosaccharid interactions. Blocking the adhesion would be a way to fight against the infection. The anti-adhesion therapy targets several bacterial lectins involved in adhesiveness and biofilm formation. In this work, the recently identified putative fimbrial lectins CupB6 and CupE6 have been studied. Expression and purification tests followed by crystallization trials have been performed. In parallel, attempts to identify the natural ligand of CupB6 were also carried out. This work also presents a systematic characterization of the inhibitory effects of various galactose-derived molecules on the PA-IL lectin. Some of these molecules could be used as glycomimetic drugs thus…

Advisors/Committee Members: Varrot, Annabelle (thesis director).

Subjects/Keywords: Clonage; Expression de protéines recombinantes chez E. coli; Purification de protéines; Tests d'affinité des protéines; Résolution de stucture par des méthodes de crystallographie à rayon X; Crystallogénèse; Cloning; Expression of recombinant proteins; Protein purification; Specificity determination; 3D structure determination by X-ray crystallography; Crystallogenesis

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Ganne, G. (2013). Etudes structurales et fonctionnelles de lectines et d'adhésines chez Pseudomonas aeruginosa : Structural and functional characterisation of lectines and adhesins from Pseudomonas aeruginosa. (Doctoral Dissertation). Université de Grenoble. Retrieved from http://www.theses.fr/2013GRENV005

Chicago Manual of Style (16^th Edition):

Ganne, Géraldine. “Etudes structurales et fonctionnelles de lectines et d'adhésines chez Pseudomonas aeruginosa : Structural and functional characterisation of lectines and adhesins from Pseudomonas aeruginosa.” 2013. Doctoral Dissertation, Université de Grenoble. Accessed February 28, 2021. http://www.theses.fr/2013GRENV005.

MLA Handbook (7^th Edition):

Ganne, Géraldine. “Etudes structurales et fonctionnelles de lectines et d'adhésines chez Pseudomonas aeruginosa : Structural and functional characterisation of lectines and adhesins from Pseudomonas aeruginosa.” 2013. Web. 28 Feb 2021.

Vancouver:

Ganne G. Etudes structurales et fonctionnelles de lectines et d'adhésines chez Pseudomonas aeruginosa : Structural and functional characterisation of lectines and adhesins from Pseudomonas aeruginosa. [Internet] [Doctoral dissertation]. Université de Grenoble; 2013. [cited 2021 Feb 28]. Available from: http://www.theses.fr/2013GRENV005.

Council of Science Editors:

Ganne G. Etudes structurales et fonctionnelles de lectines et d'adhésines chez Pseudomonas aeruginosa : Structural and functional characterisation of lectines and adhesins from Pseudomonas aeruginosa. [Doctoral Dissertation]. Université de Grenoble; 2013. Available from: http://www.theses.fr/2013GRENV005

University of Waterloo

6. Tran, Norman. Establishing Viable Systems for the Recombinant Production of Haemophilus influenzae Immunoglobulin A1 Protease.

Degree: 2020, University of Waterloo

URL: http://hdl.handle.net/10012/16146

► Immunoglobulin A1 proteases (IgAPs) are a family of enzymatic virulence factors secreted from a wide variety of pathogenic bacteria known to infect human mucosal surfaces.… (more)

Subjects/Keywords: immunoglobulin A1 proteases; protein purification; protein expression; Haemophilus influenzae

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Tran, N. (2020). Establishing Viable Systems for the Recombinant Production of Haemophilus influenzae Immunoglobulin A1 Protease. (Thesis). University of Waterloo. Retrieved from http://hdl.handle.net/10012/16146

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Tran, Norman. “Establishing Viable Systems for the Recombinant Production of Haemophilus influenzae Immunoglobulin A1 Protease.” 2020. Thesis, University of Waterloo. Accessed February 28, 2021. http://hdl.handle.net/10012/16146.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Tran, Norman. “Establishing Viable Systems for the Recombinant Production of Haemophilus influenzae Immunoglobulin A1 Protease.” 2020. Web. 28 Feb 2021.

Vancouver:

Tran N. Establishing Viable Systems for the Recombinant Production of Haemophilus influenzae Immunoglobulin A1 Protease. [Internet] [Thesis]. University of Waterloo; 2020. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/10012/16146.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Tran N. Establishing Viable Systems for the Recombinant Production of Haemophilus influenzae Immunoglobulin A1 Protease. [Thesis]. University of Waterloo; 2020. Available from: http://hdl.handle.net/10012/16146

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Northeastern University

7. Mallipeddi, Srikrishnan. Biochemical and biophysical study of cannabinoid 1 and cannabinoid 2 receptors.

Degree: PhD, School of Pharmacy, 2016, Northeastern University

URL: http://hdl.handle.net/2047/D20238132

► Cannabinoid receptors are Class A G-protein coupled receptors (GPCRs) and are involved in the modulation of a diverse set of signaling pathways in the human… (more)

▼ Cannabinoid receptors are Class A G-protein coupled receptors (GPCRs) and are involved in the modulation of a diverse set of signaling pathways in the human body. The two subtypes of cannabinoid receptors are Cannabinoid 1 receptor (CB1) and Cannabinoid 2 receptor (CB2), which are expressed predominantly in the human brain and in the immune system, respectively. Both receptors are considered to be key drug targets with enormous therapeutic potential in disease conditions ranging from neuropathic pain and neurodegenerative disorders to cardio-metabolic and inflammatory disorders. In order to develop high-affinity, selective Cannabinergic ligands with specific downstream signaling effects, we need to better understand the structural aspects of the receptor in its native environment.; To study the ligand binding site(s) of CB2, we had previously developed an experimental approach termed as Ligand Assisted Protein Structure (LAPS). Using a combination of chemically functionalized Cannabinergic ligands and site-directed mutagenesis and pharmacological studies, we were able to ascertain that AM1336, a potent CB2-selective inverse agonist, attaches covalently to the receptor through critical interactions with the two cysteines on transmembrane helix 7 of CB2. To confirm directly the covalent attachment of AM1336 to amino acid(s) on CB2, we used a bottom-up proteomic approach using mass spectrometry. We used a baculovirus-based insect cell expression system to express and purify functional, full-length human CB2 receptor for mass spectrometric studies. Purified CB2 was treated with AM1336 and analyzed using bottom-up proteomic analysis to identify the modified amino acid residue(s). Using high resolution mass spectrometric analysis, we were able to confirm the covalent modification of C284 (7.38) of transmembrane helix 7, confirming the previous biochemical evidence. Using comprehensive modeling and docking techniques, the binding site of AM1336 was visualized and other receptor-ligand interactions were identified.; From previous reports on other GPCRs, following ligand binding, the receptor undergoes critical conformational changes, which determine the activation of various downstream signaling pathways. In order to study the receptor dynamics of CB2 in its native lipid environment, we developed a system for future structural studies. We incorporated purified functional CB2 into lipid bilayer mimics known as nanodiscs and further purified CB2-incorporated nanodiscs using size-exclusion and affinity chromatography. This work could provide us a system to study the changes in the structural conformation of CB2, in the presence of cannabinergic ligands, using hydrogen-deuterium exchange mass spectrometry. This system would not only enable us to determine the key conformational changes in CB2, following ligand binding, but also provide critical insight into the structural motifs involved in receptor function and downstream signaling pathway activation.; The LAPS approach-based study of CB1 has proven to be a difficult challenge…

Subjects/Keywords: cannabinoid; GPCR; mass spectrometry; protein expression; protein purification

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Mallipeddi, S. (2016). Biochemical and biophysical study of cannabinoid 1 and cannabinoid 2 receptors. (Doctoral Dissertation). Northeastern University. Retrieved from http://hdl.handle.net/2047/D20238132

Chicago Manual of Style (16^th Edition):

Mallipeddi, Srikrishnan. “Biochemical and biophysical study of cannabinoid 1 and cannabinoid 2 receptors.” 2016. Doctoral Dissertation, Northeastern University. Accessed February 28, 2021. http://hdl.handle.net/2047/D20238132.

MLA Handbook (7^th Edition):

Mallipeddi, Srikrishnan. “Biochemical and biophysical study of cannabinoid 1 and cannabinoid 2 receptors.” 2016. Web. 28 Feb 2021.

Vancouver:

Mallipeddi S. Biochemical and biophysical study of cannabinoid 1 and cannabinoid 2 receptors. [Internet] [Doctoral dissertation]. Northeastern University; 2016. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/2047/D20238132.

Council of Science Editors:

Mallipeddi S. Biochemical and biophysical study of cannabinoid 1 and cannabinoid 2 receptors. [Doctoral Dissertation]. Northeastern University; 2016. Available from: http://hdl.handle.net/2047/D20238132

8. Österberg, Emmy. Ro52 : Structure and interactions of constructs of RING and B-box.

Degree: The Institute of Technology, 2014, Linköping UniversityLinköping University

URL: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110362

► The ubiquitination process is vital to maintain the protein homeostasis in the cell. With high specificity it regulates degradation of proteins by tagging them… (more)

Subjects/Keywords: Structural biology; Ro52; TRIM21; Ubiquitination; Western blot; Protein expression; Protein purification

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APA (6^th Edition):

Österberg, E. (2014). Ro52 : Structure and interactions of constructs of RING and B-box. (Thesis). Linköping UniversityLinköping University. Retrieved from http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110362

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Österberg, Emmy. “Ro52 : Structure and interactions of constructs of RING and B-box.” 2014. Thesis, Linköping UniversityLinköping University. Accessed February 28, 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110362.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Österberg, Emmy. “Ro52 : Structure and interactions of constructs of RING and B-box.” 2014. Web. 28 Feb 2021.

Vancouver:

Österberg E. Ro52 : Structure and interactions of constructs of RING and B-box. [Internet] [Thesis]. Linköping UniversityLinköping University; 2014. [cited 2021 Feb 28]. Available from: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110362.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Österberg E. Ro52 : Structure and interactions of constructs of RING and B-box. [Thesis]. Linköping UniversityLinköping University; 2014. Available from: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110362

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Victoria University of Wellington

9. Trounson, Vaughan. Identifying Human Host Cell Protein Targets of the Bartonella Effector Protein (Bep) Fic Domains.

Degree: 2016, Victoria University of Wellington

URL: http://hdl.handle.net/10063/5286

► The genus Bartonellae represents an increasing number of emerging bacterial pathogens that utilises an unusual infection strategy, parasitising the red blood cells of their mammalian… (more)

▼ The genus Bartonellae represents an increasing number of emerging bacterial pathogens that utilises an unusual infection strategy, parasitising the red blood cells of their mammalian host. The most common species to infect humans are B. henselae and B. quintana. B. henselae is transmitted between cats by the cat flea, although occasionally infects humans via cat scratches or bites, causing cat-scratch disease (CSD). CSD is characterised by enlarged tender lymph nodes and fever. B. henselae also infects the endothelial cells of both its hosts; likely a factor in disease progression. B. quintana, the cause of trench fever during WWI, is spread people by the body louse. Trench fever is characterised by relapsing fever, headache, and bone pain. B. quintana is also able to infect human endothelial cells. These bacteria secrete a range of Bartonella effector proteins (Beps) via a Type IV secretion system, directly into endothelial cells, subverting host cell processes and resulting in internalisation of the bacteria. Beps have a range of functions, many of which are not fully characterised. B. henselae secretes three Beps (BepA-C) that contain a filamentation induced by cAMP (Fic) domain and a Bartonella Intracellular Delivery (BID) domain, with BepA being the best studied. BepA’s BID domain is responsible for intracellular delivery as well as inhibition of apoptosis by the host cell, however the exact function of the Fic domain remains unknown. Fic-containing bacterial toxins catalyse the transfer of an AMP moiety from ATP onto a host cell protein. This AMPylation frequently inactivates these proteins resulting in disrupted host cell processes and cytotoxicity. BepA has previously been shown to possess AMPylation activity, although the host target protein(s) are unknown. Evidence suggests that these proteins are novel targets. The aim of this study was to create protein constructs containing these Fic domains, and to develop techniques to identify the host cell target proteins post AMPylation. To this end, both a fluorescent ATP analogue and a fluorescent click chemistry based approach were utilised. While no target protein was identified, a basic methodology was developed for protein production and target protein identification that could be further developed. Advisors/Committee Members: Mackichan, Joanna.

Subjects/Keywords: Bartonella henselae; AMPylation; Protein Purification; Adenylylation; Fic Domain; Protein expression

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Trounson, V. (2016). Identifying Human Host Cell Protein Targets of the Bartonella Effector Protein (Bep) Fic Domains. (Masters Thesis). Victoria University of Wellington. Retrieved from http://hdl.handle.net/10063/5286

Chicago Manual of Style (16^th Edition):

Trounson, Vaughan. “Identifying Human Host Cell Protein Targets of the Bartonella Effector Protein (Bep) Fic Domains.” 2016. Masters Thesis, Victoria University of Wellington. Accessed February 28, 2021. http://hdl.handle.net/10063/5286.

MLA Handbook (7^th Edition):

Trounson, Vaughan. “Identifying Human Host Cell Protein Targets of the Bartonella Effector Protein (Bep) Fic Domains.” 2016. Web. 28 Feb 2021.

Vancouver:

Trounson V. Identifying Human Host Cell Protein Targets of the Bartonella Effector Protein (Bep) Fic Domains. [Internet] [Masters thesis]. Victoria University of Wellington; 2016. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/10063/5286.

Council of Science Editors:

Trounson V. Identifying Human Host Cell Protein Targets of the Bartonella Effector Protein (Bep) Fic Domains. [Masters Thesis]. Victoria University of Wellington; 2016. Available from: http://hdl.handle.net/10063/5286

University of Oxford

10. Ossa, Felipe. Regulating proteostasis : a study of interactions between HSP70 proteins, HSP40 proteins, and the sigma 1 receptor.

Degree: PhD, 2018, University of Oxford

URL: http://ora.ox.ac.uk/objects/uuid:8a679086-9fb4-4c7d-bfb4-45d7271f86c2 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.770626

► Proteostasis encompasses a number of cellular processes that are essential for maintaining cellular protein machinery, ranging from synthesis and folding through to degradation, prevention of… (more)

▼ Proteostasis encompasses a number of cellular processes that are essential for maintaining cellular protein machinery, ranging from synthesis and folding through to degradation, prevention of aggregation and even the mediation of constitutive function. This fundamental cellular process is linked to several clinically important phenomena, in particular neurodegeneration, cancer and ageing. Understanding the proteostatic machinery requires not just an individual examination of the key players in the system, namely the protein chaperones, but also an examination of the vast network of regulatory machinery that underpins their function. The work presented here examines two types of regulators: the HSP40 proteins and the Sigma 1 Receptor, in the context of their impact on the activity of HSP70 proteins, one of the key hubs of the proteostatic network. The Sigma 1 Receptor is a small (25 kDa) protein of significant clinical importance, and recent work has highlighted its links to the proteostatic machinery and its interaction with HSP70 proteins. With its unique pharmacology, Sigma 1 is now emerging as a novel ligand-operated regulator of proteostasis. Our work on the Sigma 1 Receptor was hindered by the difficulties of expressing and isolating a functional integral human membrane protein; despite extensive optimisation of the construct and procedures for expression in E. coli and reconstitution into detergent we remained unable to demonstrate functionality. The data presented here on balance argues that even under our optimised conditions the receptor structure is significantly disrupted, and that activity is largely absent. More successful studies in cellulo show the response of the oligomeric state of Sigma 1 to its ligands using a novel number and brightness-based methodology. We next turned to HSP40 regulation of HSP70s. HSP40s are canonical regulators of HSP70s and act by stimulating ATPase activity using J-domains. We hypothesised that because of their different roles in the cell, the human cytosolic HSP70 and HSC70 may preferentially interact with or be stimulated by J-domains from different HSP40s. Using the J-domains of DNAJB2, DNAJB6, and DNAJC5, we show that the rank order of ATPase stimulation is preserved for HSP70 and HSC70, but that the stimulated activity of HSP70 is less than that of HSC70. Furthermore, we present data indicating that the HSP70 C-terminal domain plays a role in determining the response of HSC70 and HSP70 to regulation by HSP40 co-chaperone J-domains. Finally, we confirm the importance of the G/F rich region for the structure and function of a B class J-domain from humans, DNAJB6.

Subjects/Keywords: Protein Expression and Purification; NMR; Protein-Protein Interactions; Biophysics; Protein Chaperones; Fluorescence Microscopy

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Ossa, F. (2018). Regulating proteostasis : a study of interactions between HSP70 proteins, HSP40 proteins, and the sigma 1 receptor. (Doctoral Dissertation). University of Oxford. Retrieved from http://ora.ox.ac.uk/objects/uuid:8a679086-9fb4-4c7d-bfb4-45d7271f86c2 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.770626

Chicago Manual of Style (16^th Edition):

Ossa, Felipe. “Regulating proteostasis : a study of interactions between HSP70 proteins, HSP40 proteins, and the sigma 1 receptor.” 2018. Doctoral Dissertation, University of Oxford. Accessed February 28, 2021. http://ora.ox.ac.uk/objects/uuid:8a679086-9fb4-4c7d-bfb4-45d7271f86c2 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.770626.

MLA Handbook (7^th Edition):

Ossa, Felipe. “Regulating proteostasis : a study of interactions between HSP70 proteins, HSP40 proteins, and the sigma 1 receptor.” 2018. Web. 28 Feb 2021.

Vancouver:

Ossa F. Regulating proteostasis : a study of interactions between HSP70 proteins, HSP40 proteins, and the sigma 1 receptor. [Internet] [Doctoral dissertation]. University of Oxford; 2018. [cited 2021 Feb 28]. Available from: http://ora.ox.ac.uk/objects/uuid:8a679086-9fb4-4c7d-bfb4-45d7271f86c2 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.770626.

Council of Science Editors:

Ossa F. Regulating proteostasis : a study of interactions between HSP70 proteins, HSP40 proteins, and the sigma 1 receptor. [Doctoral Dissertation]. University of Oxford; 2018. Available from: http://ora.ox.ac.uk/objects/uuid:8a679086-9fb4-4c7d-bfb4-45d7271f86c2 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.770626

Virginia Tech

11. Liu, Jianyun. Plant-derived Murine IL-12 and Ricin B-Murine IL-12 Fusions.

Degree: PhD, Plant Pathology, Physiology, and Weed Science, 2006, Virginia Tech

URL: http://hdl.handle.net/10919/30190

► Interleukin-12 (IL-12), an important immuno-modulator for cell-mediated immunity, shows significant potential as a vaccine adjuvant and anti-cancer therapeutic. However, its clinical application is limited by… (more)

▼ Interleukin-12 (IL-12), an important immuno-modulator for cell-mediated immunity, shows significant potential as a vaccine adjuvant and anti-cancer therapeutic. However, its clinical application is limited by lack of an effective bioproduction system and by toxicity associated with systemic administration of IL-12. The goals of this research were to determine whether plants can serve as an effective production system for bioactive IL-12, a complex 70kDa glycoprotein cytokine, and whether the plant lectin RTB can facilitate mucosal delivery of IL-12 to immune responsive sites. Transgenic tobacco plants expressing murine IL-12 were generated and characterized. To ensure stochiometric expression of the two separately encoded, disulfide-linked subunits of IL-12 (p35 and p40), a single-chain form of mouse IL-12 (mIL-12) was utilized. Hairy root cultures, as a fast-growing bioproduction system were developed from high expressers of mIL-12. A purification scheme was developed to purify plant-derived mIL-12 from hairy roots and purified mIL-12 was used to assess IL-12 bioactivity in vitro in mouse splenocytes and in vivo in mouse intranasal vaccination trials. Plant-derived mIL-12 triggered induction of interferon-gamma secretion from mouse splenocytes as well as stimulation of cell proliferation with comparable activities to those observed for the animal-cell-derived mIL-12. Mouse vaccination trials using GFP as the antigen and CT as the adjuvant suggested that plant-derived mIL-12 enhanced Th1 immunity and exhibited similar activity to animal-cell-derived mIL-12 in vivo. Plant-derived IL-12 itself was non-immunogenic suggesting conformational equivalency to endogenous mouse IL-12. Ricin B (RTB), the non-toxic carbohydrate-binding subunit of ricin, directs uptake of ricin into mammalian cells and the intracellular trafficking of ricin A, the catalytic subunit of ricin. RTB's function suggests that it may work as a molecular carrier for effective mucosal delivery of IL-12. To prove this hypothesis, transgenic plants producing RTB:IL-12 fusions were generated and characterized. Our results demonstrated that RTB fused to the carboxyl-terminus of IL-12 maintained full lectin activity and IL-12 bioactivity. RTB fused to the amino-terminus of IL-12 did not show lectin activity due to steric hinderance. Purified IL-12:RTB from transgenic plant tissue was tested in an in vitro mucosal-associated lymphoid tissue (MALT) assay. The results indicate that RTB facilitates the binding of IL-12 to the epithelial cells and presentation of IL-12 to immune responsive cells. In conclusion, my research has shown that transgenic plants are capable of producing valuable bioactive proteins, such as IL-12. Plant-derived mIL-12 exhibited similar activity to animal-cell-derived mIL-12 both in vitro and in vivo. Fusion of IL-12 with the RTB lectin facilitates the delivery of IL-12 to mucosal immune responsive cells and thus may serve as a molecular carrier to enhance IL-12 efficacy and reduce the side-effects associated with systemic… Advisors/Committee Members: Cramer, Carole L. (committeechair), He, Yongqun (committee member), Dolbin-MacNab, Megan L. (committee member), Gogal, Robert M. Jr. (committee member), Nessler, Craig L. (committeecochair).

Subjects/Keywords: adjuvant; recombinant expression; lectin; interleukin-12; glycoprotein; vaccine; plant bioproduction system; single-chain protein; protein purification

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APA (6^th Edition):

Liu, J. (2006). Plant-derived Murine IL-12 and Ricin B-Murine IL-12 Fusions. (Doctoral Dissertation). Virginia Tech. Retrieved from http://hdl.handle.net/10919/30190

Chicago Manual of Style (16^th Edition):

Liu, Jianyun. “Plant-derived Murine IL-12 and Ricin B-Murine IL-12 Fusions.” 2006. Doctoral Dissertation, Virginia Tech. Accessed February 28, 2021. http://hdl.handle.net/10919/30190.

MLA Handbook (7^th Edition):

Liu, Jianyun. “Plant-derived Murine IL-12 and Ricin B-Murine IL-12 Fusions.” 2006. Web. 28 Feb 2021.

Vancouver:

Liu J. Plant-derived Murine IL-12 and Ricin B-Murine IL-12 Fusions. [Internet] [Doctoral dissertation]. Virginia Tech; 2006. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/10919/30190.

Council of Science Editors:

Liu J. Plant-derived Murine IL-12 and Ricin B-Murine IL-12 Fusions. [Doctoral Dissertation]. Virginia Tech; 2006. Available from: http://hdl.handle.net/10919/30190

12. Rashev, Margarita. Optimizing the large-scale production of Saw1 and the Saw1-Rad1-Rad10 nuclease complex for structural studies.

Degree: MSc, 2017, McMaster University

URL: http://hdl.handle.net/11375/21472

►

Yeast Rad1-Rad10 is a structure specific nuclease that processes branched double-strand break (DSB) repair intermediates; the persistence of which can impede normal DNA metabolism. The… (more)

Subjects/Keywords: DNA repair; Recombinant protein expression and purification; Single strand annealing; structure selective nuclease

…Escherichia coli, Protein Expression and Purification (2017), doi: 10.1016/j.pep… …Recombinant Saw1 is mostly insoluble when expressed in E. coli expression cell lines Effect of cell… …DECLARATION OF ACADEMIC ACHIEVEMENT I optimized the expression and purification of Saccharomyces… …for each homolog. I optimized the expression and purification of Saccharomyces cerevisiae… …step Purification of His-Saw1…

10 more images…

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APA (6^th Edition):

Rashev, M. (2017). Optimizing the large-scale production of Saw1 and the Saw1-Rad1-Rad10 nuclease complex for structural studies. (Masters Thesis). McMaster University. Retrieved from http://hdl.handle.net/11375/21472

Chicago Manual of Style (16^th Edition):

Rashev, Margarita. “Optimizing the large-scale production of Saw1 and the Saw1-Rad1-Rad10 nuclease complex for structural studies.” 2017. Masters Thesis, McMaster University. Accessed February 28, 2021. http://hdl.handle.net/11375/21472.

MLA Handbook (7^th Edition):

Rashev, Margarita. “Optimizing the large-scale production of Saw1 and the Saw1-Rad1-Rad10 nuclease complex for structural studies.” 2017. Web. 28 Feb 2021.

Vancouver:

Rashev M. Optimizing the large-scale production of Saw1 and the Saw1-Rad1-Rad10 nuclease complex for structural studies. [Internet] [Masters thesis]. McMaster University; 2017. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/11375/21472.

Council of Science Editors:

Rashev M. Optimizing the large-scale production of Saw1 and the Saw1-Rad1-Rad10 nuclease complex for structural studies. [Masters Thesis]. McMaster University; 2017. Available from: http://hdl.handle.net/11375/21472

University of Manchester

13. Alzahrani, Ateeq Ahmed h. Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance.

Degree: 2012, University of Manchester

URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:180804

►

The cystic fibrosis transmembrane conductance regulator (CFTR) is a transmembrane protein that functions as an ion channel. Mutations in this protein cause Cystic Fibrosis. For… (more)

▼

The cystic fibrosis transmembrane conductance regulator (CFTR) is a transmembrane protein that functions as an ion channel. Mutations in this protein cause Cystic Fibrosis. For this reason, it is important to study the structure and function of CFTR. In this study, constructs of CFTR (C-terminii), a CFTR-interacting protein and full-length CFTR were cloned, expressed and purified for structural and functional studies. The purified C-terminal polypeptides of CFTR were soluble and shown to interact with NHERF1 PDZ 1 (a CFTR-interacting protein). The CFTR C-terminus and NHERF1 PDZ 1 domain were co-expressed and co-purified. The purified complex showed a strong interaction that might induces a conformational change. Site-directed mutation of the C-terminus of CFTR was performed in order to examine the effect of removing a potentially flexible amino acid (Arginine) on protein crystallization. Pull-down assay experiments with full-length CFTR demonstrated an interaction between CFTR (in DDM detergent) and NHERF1 PDZ 1(+). No interaction was observed for CFTR in LPG (a relatively denaturing detergent) and NHERF1, implying that the interaction between the PDZ motive of CFTR and NHERF1 requires a stable folded structure for both proteins. In addition, full-length CFTR in DDM has been studied by electron microscopy and Single Particle Analysis in the presence of NHERF1 PDZ 1(+). A 3D structure was generated for the CFTR-NHERF1 PDZ 1(+) complex at a resolution of ~ 18 Å. This 3D structure showed a new open conformation of CFTR (V shape). In comparable studies with CFTR alone, a 3D structure was generated at a resolution of 27 Å and this structure showed a closed state as previously reported. This new data suggest a possible role for NHERF1 in terms of CFTR channel gating or activation.

Table of ContentsAcknowledgements 3Table of Contents 4List of Tables 11List of Figures 12List of Abbreviations 17Introduction 211.1 Transmembrane proteins 211.2 ABC proteins 231.3 The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) 251.3.1 CFTR-Interacting Proteins 291.3.1.1 CFTR-NHERF Interactions 301.3.1.1.1 Does NHERF-C-terminus of CFTR binding have an important role in CF? 341.4 Structural Biology of Transmembrane Proteins 351.4.1 Electron Microscpy 351.4.1.1 2D Crystals of CFTR 371.4.1.2 Single Particle Analysis of CFTR 391.4.2 The Structure of CFTR Domains 421.4.2.1 Prediction of CFTR Topology (and the location of C-terminus) 461.4.2.2 The C-terminus of CFTR 471.4.2.3 Expression and Purification of CFTR 491.5 Characterization of Protein Using Circular Dichroism 501.6 Characterization of Protein by MALDI-TOF Mass Spectrometry 541.7 Aims and Objectives of this Study 56Materials and Methods 582.1 Materials 582.1.1 Source of the CFTR Protein 582.1.2 Growth Media 582.1.3 Antibiotics 582.1.4 Inducers 592.1.5 Buffers and Solutions 592.1.5.1 Buffer A (Unbound Materials Washing Buffer) 592.1.5.2 Buffer B (Elution Buffer) 592.1.5.3 Buffer C (Sonication Buffer) 592.1.5.4 Dialysis Buffer 592.1.5.5 GST (the glutathione S-transferase) tag…

Advisors/Committee Members: Ford, Robert.

Subjects/Keywords: Protein expression and Purification; Electron Microscopy; Single Particle Analysis.

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Alzahrani, A. A. h. (2012). Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:180804

Chicago Manual of Style (16^th Edition):

Alzahrani, Ateeq Ahmed h. “Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance.” 2012. Doctoral Dissertation, University of Manchester. Accessed February 28, 2021. http://www.manchester.ac.uk/escholar/uk-ac-man-scw:180804.

MLA Handbook (7^th Edition):

Alzahrani, Ateeq Ahmed h. “Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance.” 2012. Web. 28 Feb 2021.

Vancouver:

Alzahrani AAh. Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance. [Internet] [Doctoral dissertation]. University of Manchester; 2012. [cited 2021 Feb 28]. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:180804.

Council of Science Editors:

Alzahrani AAh. Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance. [Doctoral Dissertation]. University of Manchester; 2012. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:180804

University of Manchester

14. Alzahrani, Ateeq Ahmed Hassan. Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance.

Degree: PhD, 2012, University of Manchester

URL: https://www.research.manchester.ac.uk/portal/en/theses/structural-biology-of-cystic-fibrosis-transmembrane-conductance-regulator-an-atpbinding-cassette-protein-of-medical-importance(b8d020d3-24d7-474a-afb5-a112e38ac027).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.606912

► The cystic fibrosis transmembrane conductance regulator (CFTR) is a transmembrane protein that functions as an ion channel. Mutations in this protein cause Cystic Fibrosis. For… (more)

Subjects/Keywords: 616.3; Protein expression and Purification; Electron Microscopy; Single Particle Analysis

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Alzahrani, A. A. H. (2012). Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance. (Doctoral Dissertation). University of Manchester. Retrieved from https://www.research.manchester.ac.uk/portal/en/theses/structural-biology-of-cystic-fibrosis-transmembrane-conductance-regulator-an-atpbinding-cassette-protein-of-medical-importance(b8d020d3-24d7-474a-afb5-a112e38ac027).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.606912

Chicago Manual of Style (16^th Edition):

Alzahrani, Ateeq Ahmed Hassan. “Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance.” 2012. Doctoral Dissertation, University of Manchester. Accessed February 28, 2021. https://www.research.manchester.ac.uk/portal/en/theses/structural-biology-of-cystic-fibrosis-transmembrane-conductance-regulator-an-atpbinding-cassette-protein-of-medical-importance(b8d020d3-24d7-474a-afb5-a112e38ac027).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.606912.

MLA Handbook (7^th Edition):

Alzahrani, Ateeq Ahmed Hassan. “Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance.” 2012. Web. 28 Feb 2021.

Vancouver:

Alzahrani AAH. Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance. [Internet] [Doctoral dissertation]. University of Manchester; 2012. [cited 2021 Feb 28]. Available from: https://www.research.manchester.ac.uk/portal/en/theses/structural-biology-of-cystic-fibrosis-transmembrane-conductance-regulator-an-atpbinding-cassette-protein-of-medical-importance(b8d020d3-24d7-474a-afb5-a112e38ac027).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.606912.

Council of Science Editors:

Alzahrani AAH. Structural biology of Cystic Fibrosis Transmembrane Conductance Regulator, an ATP-binding cassette protein of medical importance. [Doctoral Dissertation]. University of Manchester; 2012. Available from: https://www.research.manchester.ac.uk/portal/en/theses/structural-biology-of-cystic-fibrosis-transmembrane-conductance-regulator-an-atpbinding-cassette-protein-of-medical-importance(b8d020d3-24d7-474a-afb5-a112e38ac027).html ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.606912

Iowa State University

15. Kumar, Nitin. Structure, function, and regulation of bacterial efflux pumps.

Degree: 2016, Iowa State University

URL: https://lib.dr.iastate.edu/etd/15985

► The continued inappropriate use of antibiotics over the years has led to the development of resistance mechanisms in microorganisms against multiple drugs. One of the… (more)

Subjects/Keywords: Cloning; Crystallization; Expression; Protein; Purification; X-ray; Chemistry

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APA (6^th Edition):

Kumar, N. (2016). Structure, function, and regulation of bacterial efflux pumps. (Thesis). Iowa State University. Retrieved from https://lib.dr.iastate.edu/etd/15985

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Kumar, Nitin. “Structure, function, and regulation of bacterial efflux pumps.” 2016. Thesis, Iowa State University. Accessed February 28, 2021. https://lib.dr.iastate.edu/etd/15985.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Kumar, Nitin. “Structure, function, and regulation of bacterial efflux pumps.” 2016. Web. 28 Feb 2021.

Vancouver:

Kumar N. Structure, function, and regulation of bacterial efflux pumps. [Internet] [Thesis]. Iowa State University; 2016. [cited 2021 Feb 28]. Available from: https://lib.dr.iastate.edu/etd/15985.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Kumar N. Structure, function, and regulation of bacterial efflux pumps. [Thesis]. Iowa State University; 2016. Available from: https://lib.dr.iastate.edu/etd/15985

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Duquesne University

16. Sparacino-Watkins, Courtney. Nitrate metabolism in the Epsilonproteobacteria: Campylobacter jejuni and Sulfurospirillum barnesii.

Degree: PhD, Chemistry and Biochemistry, 2011, Duquesne University

URL: https://dsc.duq.edu/etd/1227

► In this study the nitrate metabolism of Campylobacter jejuni and Sulfurospirillum barnesii will be examined, specifically the periplasmic nitrate reductase (Nap) enzyme which is transforms… (more)

Subjects/Keywords: Campylobacter; Heterologous expression; Molybdenum enzymes; Nitrate reductase; Periplasmic; Protein purification

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APA (6^th Edition):

Sparacino-Watkins, C. (2011). Nitrate metabolism in the Epsilonproteobacteria: Campylobacter jejuni and Sulfurospirillum barnesii. (Doctoral Dissertation). Duquesne University. Retrieved from https://dsc.duq.edu/etd/1227

Chicago Manual of Style (16^th Edition):

Sparacino-Watkins, Courtney. “Nitrate metabolism in the Epsilonproteobacteria: Campylobacter jejuni and Sulfurospirillum barnesii.” 2011. Doctoral Dissertation, Duquesne University. Accessed February 28, 2021. https://dsc.duq.edu/etd/1227.

MLA Handbook (7^th Edition):

Sparacino-Watkins, Courtney. “Nitrate metabolism in the Epsilonproteobacteria: Campylobacter jejuni and Sulfurospirillum barnesii.” 2011. Web. 28 Feb 2021.

Vancouver:

Sparacino-Watkins C. Nitrate metabolism in the Epsilonproteobacteria: Campylobacter jejuni and Sulfurospirillum barnesii. [Internet] [Doctoral dissertation]. Duquesne University; 2011. [cited 2021 Feb 28]. Available from: https://dsc.duq.edu/etd/1227.

Council of Science Editors:

Sparacino-Watkins C. Nitrate metabolism in the Epsilonproteobacteria: Campylobacter jejuni and Sulfurospirillum barnesii. [Doctoral Dissertation]. Duquesne University; 2011. Available from: https://dsc.duq.edu/etd/1227

University of South Africa

17. Kgokolo, Samuel Maphalle. Optimization of purification and characterisation of over-expressed rotavirus capsid protein VP6 .

Degree: 2017, University of South Africa

URL: http://hdl.handle.net/10500/23726

► Rotavirus is responsible for the death of many children annually, and current vaccines have lower efficiency in developing countries. A reverse translated consensus gene sequence… (more)

Subjects/Keywords: Chromatography; Circular dichroism; Escherichia coli; Fluorescence; Plasmid; Protein conformation; Protein expression; Purification; Rotavirus; VP6

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APA (6^th Edition):

Kgokolo, S. M. (2017). Optimization of purification and characterisation of over-expressed rotavirus capsid protein VP6 . (Masters Thesis). University of South Africa. Retrieved from http://hdl.handle.net/10500/23726

Chicago Manual of Style (16^th Edition):

Kgokolo, Samuel Maphalle. “Optimization of purification and characterisation of over-expressed rotavirus capsid protein VP6 .” 2017. Masters Thesis, University of South Africa. Accessed February 28, 2021. http://hdl.handle.net/10500/23726.

MLA Handbook (7^th Edition):

Kgokolo, Samuel Maphalle. “Optimization of purification and characterisation of over-expressed rotavirus capsid protein VP6 .” 2017. Web. 28 Feb 2021.

Vancouver:

Kgokolo SM. Optimization of purification and characterisation of over-expressed rotavirus capsid protein VP6 . [Internet] [Masters thesis]. University of South Africa; 2017. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/10500/23726.

Council of Science Editors:

Kgokolo SM. Optimization of purification and characterisation of over-expressed rotavirus capsid protein VP6 . [Masters Thesis]. University of South Africa; 2017. Available from: http://hdl.handle.net/10500/23726

Arizona State University

18. Deb, Arpan. Towards a Structural and Functional Insight into the Human Immunodeficiency Virus (HIV) – 1 Membrane Protein, Vpu.

Degree: Molecular and Cellular Biology, 2016, Arizona State University

URL: http://repository.asu.edu/items/40285

► Viral protein U (Vpu) is a type-III integral membrane protein encoded by the Human Immunodeficiency Virus-1 (HIV- 1). It is expressed in infected host cells… (more)

Subjects/Keywords: Molecular biology; Biophysics; Biochemistry; Crystallography; HIV; Membrane Protein Expression; Membrane Protein Purification; NMR Spectroscopy; Vpu

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APA (6^th Edition):

Deb, A. (2016). Towards a Structural and Functional Insight into the Human Immunodeficiency Virus (HIV) – 1 Membrane Protein, Vpu. (Doctoral Dissertation). Arizona State University. Retrieved from http://repository.asu.edu/items/40285

Chicago Manual of Style (16^th Edition):

Deb, Arpan. “Towards a Structural and Functional Insight into the Human Immunodeficiency Virus (HIV) – 1 Membrane Protein, Vpu.” 2016. Doctoral Dissertation, Arizona State University. Accessed February 28, 2021. http://repository.asu.edu/items/40285.

MLA Handbook (7^th Edition):

Deb, Arpan. “Towards a Structural and Functional Insight into the Human Immunodeficiency Virus (HIV) – 1 Membrane Protein, Vpu.” 2016. Web. 28 Feb 2021.

Vancouver:

Deb A. Towards a Structural and Functional Insight into the Human Immunodeficiency Virus (HIV) – 1 Membrane Protein, Vpu. [Internet] [Doctoral dissertation]. Arizona State University; 2016. [cited 2021 Feb 28]. Available from: http://repository.asu.edu/items/40285.

Council of Science Editors:

Deb A. Towards a Structural and Functional Insight into the Human Immunodeficiency Virus (HIV) – 1 Membrane Protein, Vpu. [Doctoral Dissertation]. Arizona State University; 2016. Available from: http://repository.asu.edu/items/40285

University of Southern California

19. Ma, Tao. Expression and purification of different elastin like polypeptides (ELPs) constructs for therapeutic applications.

Degree: MS, Pharmaceutical Sciences, 2015, University of Southern California

URL: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/592829/rec/2687

► There are many rationales for modifying recombinant proteins by using polymers, which include: i) altering permeability/diffusion via controlling their hydrodynamic radius; ii) modulating mediators of… (more)

▼ There are many rationales for modifying recombinant proteins by using polymers, which include: i) altering permeability/diffusion via controlling their hydrodynamic radius; ii) modulating mediators of controlled release; and iii) improving their avidity to molecular/cellular targets. Many of these strategies require covalent bioconjugation, which are inefficient and may compromise protein activity. An emerging solution to this challenge is to develop gene products that act like polymers. One such polymer is known as the Elastin Like Polypeptides (ELPs). ELPs are ‘protein polymers’ composed of repetitive amino acid sequence derived from the human extracellular matrix protein called tropoelastin. In addition to their high molecular weight, which exceeds 80 kDa, ELPs undergo thermo‐responsive phase separation. ELPs are highly soluble in aqueous solutions below their transition temperature (Tt) and rapidly self‐assemble into viscous coacervate particles above Tt. Tt can be modulated by changing the molecular weight and hydrophobicity of the ELP. In this thesis, ELPs are developed as a carrier for lacritin, which is a novel tear glycoprotein that promotes tear secretion, maintains ocular surface integrity and reduces formation of lymphocytic foci in lacrimal gland. Free lacritin increases the basal tear secretion in rabbits; however, it must be administered frequently. The rationale for exploring fusions of lacritin and ELP is that they may prolong the retention on the anterior segment of the eye. In order to explore this strategy, my thesis has focused on scaling‐up the production of high purity, low endotoxin level material for subsequent evaluation treating in models of aqueous‐deficient dry eye diseases. E. coli was used for expression of the lacritin‐ELP fusion construct and size exclusion chromatography (SEC) was used to separate LV96 from cleavage product. Endotoxin removal was explored using polymyxin chromatography. This thesis represents the first report of scaling‐up production of low endotoxin level lacritin ELP fusions. Advisors/Committee Members: Hamm-Alvarez, Sarah F. (Committee Chair), Mackay, John Andrew (Committee Member), Shen, Wei-Chiang (Committee Member).

Subjects/Keywords: elastin like polypeptides; ELPs; lacritin; protein expression; scaling-up of protein purification; endotoxin removal

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APA (6^th Edition):

Ma, T. (2015). Expression and purification of different elastin like polypeptides (ELPs) constructs for therapeutic applications. (Masters Thesis). University of Southern California. Retrieved from http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/592829/rec/2687

Chicago Manual of Style (16^th Edition):

Ma, Tao. “Expression and purification of different elastin like polypeptides (ELPs) constructs for therapeutic applications.” 2015. Masters Thesis, University of Southern California. Accessed February 28, 2021. http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/592829/rec/2687.

MLA Handbook (7^th Edition):

Ma, Tao. “Expression and purification of different elastin like polypeptides (ELPs) constructs for therapeutic applications.” 2015. Web. 28 Feb 2021.

Vancouver:

Ma T. Expression and purification of different elastin like polypeptides (ELPs) constructs for therapeutic applications. [Internet] [Masters thesis]. University of Southern California; 2015. [cited 2021 Feb 28]. Available from: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/592829/rec/2687.

Council of Science Editors:

Ma T. Expression and purification of different elastin like polypeptides (ELPs) constructs for therapeutic applications. [Masters Thesis]. University of Southern California; 2015. Available from: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/592829/rec/2687

University of Saskatchewan

20. Nambisan, Anand Krishnan. Biochemical characterization of the psychiatric risk protein, Disrupted in Schizophrenia 1 (DISC1).

Degree: 2020, University of Saskatchewan

URL: http://hdl.handle.net/10388/13013

► Disrupted in Schizophrenia 1 (DISC1) is a psychiatric disease risk gene implicated in numerous mental disorders including schizophrenia. Many isoforms of DISC1 are found in… (more)

Subjects/Keywords: DISC1; GSK3β; Neuroscience; Neuropsychiatric disorders Protein biochemistry; Protein expression and purification; Structural investigation

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APA (6^th Edition):

Nambisan, A. K. (2020). Biochemical characterization of the psychiatric risk protein, Disrupted in Schizophrenia 1 (DISC1). (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/13013

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Nambisan, Anand Krishnan. “Biochemical characterization of the psychiatric risk protein, Disrupted in Schizophrenia 1 (DISC1).” 2020. Thesis, University of Saskatchewan. Accessed February 28, 2021. http://hdl.handle.net/10388/13013.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Nambisan, Anand Krishnan. “Biochemical characterization of the psychiatric risk protein, Disrupted in Schizophrenia 1 (DISC1).” 2020. Web. 28 Feb 2021.

Vancouver:

Nambisan AK. Biochemical characterization of the psychiatric risk protein, Disrupted in Schizophrenia 1 (DISC1). [Internet] [Thesis]. University of Saskatchewan; 2020. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/10388/13013.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Nambisan AK. Biochemical characterization of the psychiatric risk protein, Disrupted in Schizophrenia 1 (DISC1). [Thesis]. University of Saskatchewan; 2020. Available from: http://hdl.handle.net/10388/13013

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Brno University of Technology

21. Bušanski, Patrik. Srovnání biotechnologických postupů přípravy čistých proteinů: Comparison of biotechnological procedures for pure proteins preparation.

Degree: 2019, Brno University of Technology

URL: http://hdl.handle.net/11012/178425

► The production of recombinant proteins is a biotechnological process during which proteins are produced in foreign organisms by gen manipulation. To form a recombinant plasmid… (more)

Subjects/Keywords: rekombinantné proteíny; expresné systémy; izolácia a purifikácia proteínov; recombinant proteins; expression systems; isolation and purification of proteins

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APA (6^th Edition):

Bušanski, P. (2019). Srovnání biotechnologických postupů přípravy čistých proteinů: Comparison of biotechnological procedures for pure proteins preparation. (Thesis). Brno University of Technology. Retrieved from http://hdl.handle.net/11012/178425

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Bušanski, Patrik. “Srovnání biotechnologických postupů přípravy čistých proteinů: Comparison of biotechnological procedures for pure proteins preparation.” 2019. Thesis, Brno University of Technology. Accessed February 28, 2021. http://hdl.handle.net/11012/178425.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Bušanski, Patrik. “Srovnání biotechnologických postupů přípravy čistých proteinů: Comparison of biotechnological procedures for pure proteins preparation.” 2019. Web. 28 Feb 2021.

Vancouver:

Bušanski P. Srovnání biotechnologických postupů přípravy čistých proteinů: Comparison of biotechnological procedures for pure proteins preparation. [Internet] [Thesis]. Brno University of Technology; 2019. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/11012/178425.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Bušanski P. Srovnání biotechnologických postupů přípravy čistých proteinů: Comparison of biotechnological procedures for pure proteins preparation. [Thesis]. Brno University of Technology; 2019. Available from: http://hdl.handle.net/11012/178425

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of South Africa

22. Russell, Bonnie Leigh. Expression, solubilisation, purification and characterisation of recombinant bluetongue virus viral protein 7 .

Degree: 2018, University of South Africa

URL: http://hdl.handle.net/10500/24951

► Bluetongue virus belongs to the Orbivirus genus from the Reoviridae family. It infects predominantly domestic and wild ruminants and is economically significant worldwide. Bluetongue virus… (more)

▼ Bluetongue virus belongs to the Orbivirus genus from the Reoviridae family. It infects predominantly domestic and wild ruminants and is economically significant worldwide. Bluetongue virus VP7 forms the intercepting layer between the outer capsid (VP2 and VP5) and VP3 which surrounds the genomic material. BL21(DE3), NiCo21(DE3), C43(DE3) pLysS and KRX Escherichia coli cells were transformed with a pET28a plasmid with the cDNA sequence encoding Bluetongue virus VP7. Expression of Bluetongue virus VP7 was tested at post induction temperatures between 16˚C and 37 ˚C, at inducer concentrations between 0.1 mM and 1.0 mM isopropyl-β-D-thiogalactopyranoside in BL21(DE3), NiCo21(DE3) and C43(DE3) pLysS cells and 0.05 % and 0.15 % rhamnose for KRX cells, in two types of growth media (LB and 2xYT) and post-induction growth times between two and 16 hours. Under all conditions tested; Bluetongue virus VP7 expression was found to be predominantly in the insoluble fraction (pellet). BL21(DE3) and NiCo21(DE3) cells were chosen and grown for five hours post induction, induced with 0.1 mM isopropyl-β-D-thiogalactopyranoside and grown at a post-induction temperature of 37 ˚C. Bluetongue virus VP7 in bacterial cell inclusion bodies was solubilised using urea and a freeze-thaw step. Solubilisation was tested with urea concentrations between 2 M and 8 M, with solubilisation efficiency not increasing past 5 M urea. Solubilized Bluetongue virus VP7 was purified using nickel-affinity chromatography. Purified Bluetongue virus VP7 was then probed with far-UV circular dichroism and intrinsic fluorescence in several buffer conditions including different urea and guanidinium chloride concentrations as well as in the presence of glycerol and sodium chloride. Guanidinium chloride was able to cause Bluetongue virus VP7 unfolding, and the unfolding transition had 94 % and 89 % reversibility at 218 nm and 222 nm respectively. Bluetongue virus VP7 was shown to contain a native-like structure in 20 % glycerol and in up to 8 M urea and was found to be stable till at least 55 ˚C, even in the presence of 5 M urea. Glycerol and sodium chloride influenced the conformation of the protein resulting in different unfolding transitions. Thermal unfolding of Bluetongue virus VP7 was found to be irreversible. Advisors/Committee Members: Gildenhuys, S (advisor).

Subjects/Keywords: Bluetongue virus; Escherichia coli; Far-UV circular dichroism; Fluorescence; Inclusion bodies; Polyhistidine-tagged purification; Protein expression; Protein solubilisation; Protein stability; VP7

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APA (6^th Edition):

Russell, B. L. (2018). Expression, solubilisation, purification and characterisation of recombinant bluetongue virus viral protein 7 . (Masters Thesis). University of South Africa. Retrieved from http://hdl.handle.net/10500/24951

Chicago Manual of Style (16^th Edition):

Russell, Bonnie Leigh. “Expression, solubilisation, purification and characterisation of recombinant bluetongue virus viral protein 7 .” 2018. Masters Thesis, University of South Africa. Accessed February 28, 2021. http://hdl.handle.net/10500/24951.

MLA Handbook (7^th Edition):

Russell, Bonnie Leigh. “Expression, solubilisation, purification and characterisation of recombinant bluetongue virus viral protein 7 .” 2018. Web. 28 Feb 2021.

Vancouver:

Russell BL. Expression, solubilisation, purification and characterisation of recombinant bluetongue virus viral protein 7 . [Internet] [Masters thesis]. University of South Africa; 2018. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/10500/24951.

Council of Science Editors:

Russell BL. Expression, solubilisation, purification and characterisation of recombinant bluetongue virus viral protein 7 . [Masters Thesis]. University of South Africa; 2018. Available from: http://hdl.handle.net/10500/24951

University of Western Ontario

23. Wang, Huasheng. Structural and Functional Characterization of Non-Homologous End Joining Factors.

Degree: 2017, University of Western Ontario

URL: https://ir.lib.uwo.ca/etd/4887

► DNA double strand breaks represent the most toxic form of DNA damage. In mammals, non-homologous end-joining (NHEJ) is the primary DNA repair pathway for such… (more)

Subjects/Keywords: DNA double strand breaks; Non-homologous end joining; Protein expression; Protein purification; Protein structure; X-ray crystallography; Biochemistry

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APA (6^th Edition):

Wang, H. (2017). Structural and Functional Characterization of Non-Homologous End Joining Factors. (Thesis). University of Western Ontario. Retrieved from https://ir.lib.uwo.ca/etd/4887

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Wang, Huasheng. “Structural and Functional Characterization of Non-Homologous End Joining Factors.” 2017. Thesis, University of Western Ontario. Accessed February 28, 2021. https://ir.lib.uwo.ca/etd/4887.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Wang, Huasheng. “Structural and Functional Characterization of Non-Homologous End Joining Factors.” 2017. Web. 28 Feb 2021.

Vancouver:

Wang H. Structural and Functional Characterization of Non-Homologous End Joining Factors. [Internet] [Thesis]. University of Western Ontario; 2017. [cited 2021 Feb 28]. Available from: https://ir.lib.uwo.ca/etd/4887.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Wang H. Structural and Functional Characterization of Non-Homologous End Joining Factors. [Thesis]. University of Western Ontario; 2017. Available from: https://ir.lib.uwo.ca/etd/4887

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Alberta

24. Bonar, Pamela T. Towards a Structural Model of the Plasma Membrane Cl-/HCO3- Exchanger, AE1.

Degree: PhD, Department of Biochemistry, 2013, University of Alberta

URL: https://era.library.ualberta.ca/files/8049g639s

► AE1 is an electroneutral Cl-/HCO3- exchanger expressed in erythrocytes and the renal collecting duct. There is no high-resolution structure of the membrane domain, which alone… (more)

▼ AE1 is an electroneutral Cl-/HCO3- exchanger expressed in erythrocytes and the renal collecting duct. There is no high-resolution structure of the membrane domain, which alone is required for the transport activity of AE1. Here, a Saccharomyces cerevisiae expression and immuno-affinity purification system was developed for the AE1 membrane domain. The human AE1 membrane domain (residues 388-911), followed by a rhodopsin antibody epitope (AE1MD-Rho), was expressed at 0.3 mg/l of culture, and milligram quantities were purified to 93% homogeneity. AE1MD-Rho transport activity was indistinguishable from erythrocyte AE1, as assessed by radioactive [35S]SO42- efflux assays in reconstituted proteoliposomes. More recently, an electron microscopy structure of the AE1 membrane domain was proposed to have a similar protein fold to ClC chloride channels. A three-dimensional homology model of the AE1 membrane domain was created, using the Escherichia coli ClC channel structure as a template. This model agrees well with AE1 cysteine scanning mutagenesis data and blood group antigen sites. To investigate the transport mechanism of AE1, point mutations were introduced in regions of the AE1 homology model corresponding to sites involved in the ClC transport mechanism. The transport activity of these mutants was assessed by Cl-/HCO3- exchange assays in HEK293 cells and Xenopus laevis oocytes. Several AE1 mutations, at sites corresponding to ClC transport mechanism residues, resulted in significant changes in transport activity compared to wild-type AE1, without changes in electrogenicity or transport stoichiometry. A study of the E. coli ClC dimer interface identified tryptophan mutations, which disrupted the dimer interface of ClC. In a similar fashion, AE1 tryptophan mutations were made in an AE1 membrane domain background, using the AE1 homology model as a guide. The majority of AE1 tryptophan mutations decreased AE1 protein expression; however, no disruptions of the dimer interface were observed using chemical crosslinking. Chemical crosslinking may not have the sensitivity to monitor slight disruptions in the AE1 dimer interface, and thus alternate strategies to monitor the oligomeric state of AE1 tryptophan mutants must be investigated. Together, these studies have provided valuable insights into the structure and transport mechanism of AE1.

Subjects/Keywords: AE1 structure; homology model; Bicarbonate transport; protein expression and purification; AE1 physiology; dimer interface

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APA (6^th Edition):

Bonar, P. T. (2013). Towards a Structural Model of the Plasma Membrane Cl-/HCO3- Exchanger, AE1. (Doctoral Dissertation). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/8049g639s

Chicago Manual of Style (16^th Edition):

Bonar, Pamela T. “Towards a Structural Model of the Plasma Membrane Cl-/HCO3- Exchanger, AE1.” 2013. Doctoral Dissertation, University of Alberta. Accessed February 28, 2021. https://era.library.ualberta.ca/files/8049g639s.

MLA Handbook (7^th Edition):

Bonar, Pamela T. “Towards a Structural Model of the Plasma Membrane Cl-/HCO3- Exchanger, AE1.” 2013. Web. 28 Feb 2021.

Vancouver:

Bonar PT. Towards a Structural Model of the Plasma Membrane Cl-/HCO3- Exchanger, AE1. [Internet] [Doctoral dissertation]. University of Alberta; 2013. [cited 2021 Feb 28]. Available from: https://era.library.ualberta.ca/files/8049g639s.

Council of Science Editors:

Bonar PT. Towards a Structural Model of the Plasma Membrane Cl-/HCO3- Exchanger, AE1. [Doctoral Dissertation]. University of Alberta; 2013. Available from: https://era.library.ualberta.ca/files/8049g639s

25. Parisien, Albert. Large-Scale Production in 'Escherichia coli' TG1 and Purification of Llama Single Domain Antibody ToxA5.1 Against 'Clostridium difficile' Toxin A .

Degree: 2013, University of Ottawa

URL: http://hdl.handle.net/10393/26242

► Drug resistant strains of Clostridium difficile are a major health concern with over 3 million cases costing over 1 billion $ per year in the… (more)

▼ Drug resistant strains of Clostridium difficile are a major health concern with over 3 million cases costing over 1 billion $ per year in the United-States. The diseases associated with these bacteria (CDAD) are toxin-mediated which offers a mean of treating and lessening the severity of CDAD symptoms. Toxin inactivation via antibodies therapy can drastically reduce CDAD morbidity and this project was aiming at investigating the large-scale production and recovery of a novel llama single domain antibody (pSJF2H-ToxA5.1) in recombinant Escherichia coli TG1 targeting C. difficile enterotoxin A (TcdA). In order to achieve these objectives, the project was divided into four segments: 1) ToxA5.1 being an intracellular recombinant protein, obtaining a high biomass production was the first step towards large-scale production. To achieve HCDC, effects of initial glucose concentration and pH-stat feeding strategy were studied; 2) Upon achieving HCDC, effects of parameters such as temperature, induction timing and media supplementation with complex nitrogen sources were investigated; 3) Once large-scale production of ToxA5.1 was obtained, the recombinant protein needed to be recovered and a selective cell lysis scheme where synergistic lysis effects of Triton X-100 and temperature were studied. And finally 4) Single-step purification using nickel nanoparticles (NNP) synthesized via a modified polyol method was studied. Combining the HCDC strategy with a temperature shift and yeast extract addition at the time of induction, ToxA5.1 concentration of 127 mg/L was obtained. Synergistic and selective cell lysis using Triton X-100 and temperature was achieved where 95% of the available ToxA5.1 was recovered and still functional while ToxA5.1 fraction in the resulting lysate increased to 27% in the cell lysate. Single-step purification was achieved using the synthesized NNP which proved to be highly selective and could be used up to five times. Diameter of the NNP synthesized was controlled by using various concentration of ranging from 131 ± 80 nm to 47 ± 20 nm. Using experimental data from binding isotherm, the ToxA5.1-NNP system was modeled.

Subjects/Keywords: Single domain antibody; C. difficile toxin A; E. coli; Fermentation; pH-stat; Protein expression; Nickel nanoparticles; Protein purification; Hexahistidine-tagged recombinant protein

…of nickel nanoparticles for purification of histidine-tagged recombinant protein (… …to optimize the protein expression of the recombinant strain under HCDC conditions… …Organisms used for recombinant protein expression often possess a mechanism, which can be turned… …Temperature also influences both cell growth and recombinant protein expression in bacterial strains… …75 4.4.2 Protein expression…

10 more images…

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Parisien, A. (2013). Large-Scale Production in 'Escherichia coli' TG1 and Purification of Llama Single Domain Antibody ToxA5.1 Against 'Clostridium difficile' Toxin A . (Thesis). University of Ottawa. Retrieved from http://hdl.handle.net/10393/26242

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Parisien, Albert. “Large-Scale Production in 'Escherichia coli' TG1 and Purification of Llama Single Domain Antibody ToxA5.1 Against 'Clostridium difficile' Toxin A .” 2013. Thesis, University of Ottawa. Accessed February 28, 2021. http://hdl.handle.net/10393/26242.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Parisien, Albert. “Large-Scale Production in 'Escherichia coli' TG1 and Purification of Llama Single Domain Antibody ToxA5.1 Against 'Clostridium difficile' Toxin A .” 2013. Web. 28 Feb 2021.

Vancouver:

Parisien A. Large-Scale Production in 'Escherichia coli' TG1 and Purification of Llama Single Domain Antibody ToxA5.1 Against 'Clostridium difficile' Toxin A . [Internet] [Thesis]. University of Ottawa; 2013. [cited 2021 Feb 28]. Available from: http://hdl.handle.net/10393/26242.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Parisien A. Large-Scale Production in 'Escherichia coli' TG1 and Purification of Llama Single Domain Antibody ToxA5.1 Against 'Clostridium difficile' Toxin A . [Thesis]. University of Ottawa; 2013. Available from: http://hdl.handle.net/10393/26242

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Universidade Nova

26. Campelo, Luísa Maria Ramos. Fishing the key biological components of the bacterium Geobacter metallireducens for optimal biotechnological applications.

Degree: 2015, Universidade Nova

URL: http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/15842

►

Os resultados apresentados no capítulo 2 foram incluídos no artigo Dantas JM, Campelo LM, Duke NEC, Salgueiro CA, Pokkuluri PR (2015) "The structure of PccH… (more)

▼

Os resultados apresentados no capítulo 2 foram incluídos no artigo Dantas JM, Campelo LM, Duke NEC, Salgueiro CA, Pokkuluri PR (2015) "The structure of PccH from Geobacter sulfurreducens – a novel low reduction potential monoheme cytochrome essential for accepting electrons from an electrode", FEBS Journal, 282, 2215-2231.

The microorganims from the Geobacter genus are very interesting because of their potential use for biotechnological applications. They can transfer electrons towards extracellular terminal acceptors, which include toxic or radioactive metals. In addition, they also have the capability to convert renewable biomass into electricity. The most studied members of the Geobacter genus are the Geobacter metallireducens and Geobacter sulfurreducens bacteria. G. metallireducens displays some particular physiological aspects that are even more interesting compared to G. sulfurreducens, which include (i) their ability to reduce aromatic compounds, (ii) more efficient Fe(III) reduction rates, and (iii) the ability to use nitrate as terminal electron acceptor. Similarly, to G. sulfurreducens, a family of five homologous periplasmic triheme c-type cytochromes, designated PpcA, PpcB, PpcC, PpcE and PpcF, was identified in G. metallireducens. In G. sulfurreducens the triheme cytochromes were shown to be crucial for extracellular electron transfer and therefore it is conceivable that these proteins play a similar role in G. metallireducens. Therefore, the main goal of this work was to optimize the production of G. metallireducens triheme cytochromes as a foundation to future optimization of G. metallirudecens-based biotechnological applications. To achieve this goal, the proteins were cloned (except PpcB), heterologously expressed in Escherichia coli and purified. The preliminary characterization of PpcA and PpcF was also undertaken in the present work. The data obtained showed that both are low-spin hexacoordinated c-type cytochromes in the reduced and oxidized forms. In parallel, it was also undertaken an evolutionary study on PccH, a monoheme c-type cytochrome from G. sulfurreducens that is crucial for microbial electrosynthesis pathways in this bacterium. The results obtained suggested that PccH is a member of a new subclass within the class I cytochromes.

Advisors/Committee Members: Salgueiro, Carlos.

Subjects/Keywords: Geobacter metallireducens; c -type cytochrome; Electron transfer; Protein cloning; Heterologous expression; Protein purification; Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Química

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Campelo, L. M. R. (2015). Fishing the key biological components of the bacterium Geobacter metallireducens for optimal biotechnological applications. (Thesis). Universidade Nova. Retrieved from http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/15842

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Campelo, Luísa Maria Ramos. “Fishing the key biological components of the bacterium Geobacter metallireducens for optimal biotechnological applications.” 2015. Thesis, Universidade Nova. Accessed February 28, 2021. http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/15842.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Campelo, Luísa Maria Ramos. “Fishing the key biological components of the bacterium Geobacter metallireducens for optimal biotechnological applications.” 2015. Web. 28 Feb 2021.

Vancouver:

Campelo LMR. Fishing the key biological components of the bacterium Geobacter metallireducens for optimal biotechnological applications. [Internet] [Thesis]. Universidade Nova; 2015. [cited 2021 Feb 28]. Available from: http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/15842.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Campelo LMR. Fishing the key biological components of the bacterium Geobacter metallireducens for optimal biotechnological applications. [Thesis]. Universidade Nova; 2015. Available from: http://www.rcaap.pt/detail.jsp?id=oai:run.unl.pt:10362/15842

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Queensland University of Technology

27. Sit, Kei Chun. Investigation of the binding interactions between insulin and its receptor.

Degree: 2015, Queensland University of Technology

URL: https://eprints.qut.edu.au/87276/

► This project investigated the interactions between insulin and its receptor. A combination of computational and experimental investigations resulted in the identification of four residues in… (more)

Subjects/Keywords: insulin receptor; molecular dynamics simulations; ligand binding; principal components analysis; surface plasmon resonance; protein expression; conformational change; BIAcore; protein purification; mutagenesis

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Sit, K. C. (2015). Investigation of the binding interactions between insulin and its receptor. (Thesis). Queensland University of Technology. Retrieved from https://eprints.qut.edu.au/87276/

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Sit, Kei Chun. “Investigation of the binding interactions between insulin and its receptor.” 2015. Thesis, Queensland University of Technology. Accessed February 28, 2021. https://eprints.qut.edu.au/87276/.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Sit, Kei Chun. “Investigation of the binding interactions between insulin and its receptor.” 2015. Web. 28 Feb 2021.

Vancouver:

Sit KC. Investigation of the binding interactions between insulin and its receptor. [Internet] [Thesis]. Queensland University of Technology; 2015. [cited 2021 Feb 28]. Available from: https://eprints.qut.edu.au/87276/.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Sit KC. Investigation of the binding interactions between insulin and its receptor. [Thesis]. Queensland University of Technology; 2015. Available from: https://eprints.qut.edu.au/87276/

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Guelph

28. Neil, Kevin J. A. Biochemical Characterization of Sinapate Ester Metabolism in Arabidopsis thaliana in Response to Transient Abiotic Stress.

Degree: MS, Department of Plant Agriculture, 2015, University of Guelph

URL: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/9395

► Flavonol bisglycosides and sinapate esters are metabolites that accumulate in plants with abiotic stress. In Arabidopsis thaliana, flavonol bisglycoside levels increase during nitrogen deficiency and… (more)

Subjects/Keywords: biochemistry; secondary metabolism; Arabidopsis; phenylpropanoid; sinapate esters; stress; stress metabolism; plant molecular biology; protein expression; protein purification

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Neil, K. J. A. (2015). Biochemical Characterization of Sinapate Ester Metabolism in Arabidopsis thaliana in Response to Transient Abiotic Stress. (Masters Thesis). University of Guelph. Retrieved from https://atrium.lib.uoguelph.ca/xmlui/handle/10214/9395

Chicago Manual of Style (16^th Edition):

Neil, Kevin J A. “Biochemical Characterization of Sinapate Ester Metabolism in Arabidopsis thaliana in Response to Transient Abiotic Stress.” 2015. Masters Thesis, University of Guelph. Accessed February 28, 2021. https://atrium.lib.uoguelph.ca/xmlui/handle/10214/9395.

MLA Handbook (7^th Edition):

Neil, Kevin J A. “Biochemical Characterization of Sinapate Ester Metabolism in Arabidopsis thaliana in Response to Transient Abiotic Stress.” 2015. Web. 28 Feb 2021.

Vancouver:

Neil KJA. Biochemical Characterization of Sinapate Ester Metabolism in Arabidopsis thaliana in Response to Transient Abiotic Stress. [Internet] [Masters thesis]. University of Guelph; 2015. [cited 2021 Feb 28]. Available from: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/9395.

Council of Science Editors:

Neil KJA. Biochemical Characterization of Sinapate Ester Metabolism in Arabidopsis thaliana in Response to Transient Abiotic Stress. [Masters Thesis]. University of Guelph; 2015. Available from: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/9395

29. Lundberg, Alexander. Studying the Oligomerization of the Kinase Domain of Ephrin type-B Receptor 2 Using Analytical Ultracentrifugation and Development of a Program for Analysis of Acquired Data.

Degree: The Institute of Technology, 2014, Linköping UniversityLinköping University

URL: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110376

► Ephrin type-B receptor 2 (EphB2) is a receptor tyrosine kinase which phosphorylates proteins and thereby regulates cell migration, vascular development, axon guidance synaptic plasticity,… (more)

Subjects/Keywords: Ephrin type-B receptor 2; EphB2; Analytical Ultracentrifugation; Oligomerization; Kinase Domain; Protein Expression; Protein Purification; Programming; Analysis of Data

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Lundberg, A. (2014). Studying the Oligomerization of the Kinase Domain of Ephrin type-B Receptor 2 Using Analytical Ultracentrifugation and Development of a Program for Analysis of Acquired Data. (Thesis). Linköping UniversityLinköping University. Retrieved from http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110376

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Lundberg, Alexander. “Studying the Oligomerization of the Kinase Domain of Ephrin type-B Receptor 2 Using Analytical Ultracentrifugation and Development of a Program for Analysis of Acquired Data.” 2014. Thesis, Linköping UniversityLinköping University. Accessed February 28, 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110376.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Lundberg, Alexander. “Studying the Oligomerization of the Kinase Domain of Ephrin type-B Receptor 2 Using Analytical Ultracentrifugation and Development of a Program for Analysis of Acquired Data.” 2014. Web. 28 Feb 2021.

Vancouver:

Lundberg A. Studying the Oligomerization of the Kinase Domain of Ephrin type-B Receptor 2 Using Analytical Ultracentrifugation and Development of a Program for Analysis of Acquired Data. [Internet] [Thesis]. Linköping UniversityLinköping University; 2014. [cited 2021 Feb 28]. Available from: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110376.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Lundberg A. Studying the Oligomerization of the Kinase Domain of Ephrin type-B Receptor 2 Using Analytical Ultracentrifugation and Development of a Program for Analysis of Acquired Data. [Thesis]. Linköping UniversityLinköping University; 2014. Available from: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-110376

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

30. Boamah, Daniel. Dissecting the Histone-binding Mechanism of a PHD Finger Subtype.

Degree: PhD, Chemistry and Biochemistry, 2017, South Dakota State University

URL: https://openprairie.sdstate.edu/etd/1673

► Disordered tails of histones are critical information retrieval hub and thus, aberrations in the flow of information through these hubs are associated with a… (more)

▼ Disordered tails of histones are critical information retrieval hub and thus, aberrations in the flow of information through these hubs are associated with a number of pathological consequences in human. Mechanism for retrieval of information from these hubs is achieved by protein-protein interaction, i.e. proteins dock onto histone tails to initiate chromatin signaling. Eukaryotes have a number of small peptide binding domains that have evolved to specifically interact with histone tails, and these domains called histone readers as they read the information encoded on histone tails. Plant homeodomain (hereafter PHD) finger, a binucleated zinc finger, family is one such histone readers. Next-generation sequencing efforts on diagnosed patient’s genomes or cancer tissues show that mutations in PHD finger, particularly a subgroup of PHD fingers, are associated with number of pathological consequences. Therefore, for future understanding of the possible mechanisms for the pathological consequences, as an initial step, detailed characterization of the binding mechanism of the PHD subtype, the PHD_nW_DD, was undertaken here. Starting with human BAZ2A (bromodomain adjacent to zinc-finger 2A), one member of the PHD_nW_DD subtype that is associated with prostate cancer was utilized to probe the effect of mutations on histone tail binding. We experimentally discovered two categories of mutations that disrupt peptide binding: (1) Type-A: positions that are in contact with the peptide and (2) Type-B: positions that are remote from the peptide-binding site (distal site). For my dissertation, I focused on understanding the biochemical basis of the effects of Type-A mutations using recombinant protein chemistry and biophysical chemistry. The peptide-anchoring residue positions of BAZ2A-PHD, interestingly, are enriched in specific type of residues in a subtype specific manner. The energetics revealed that, two non-polar amino acid residues and an Aspartate residue in the treble clef knuckle make significant contributions to the formation of the hBAZ2A-histone peptide complex as mutations at these three positions completely aborted peptide binding. The energetic contributions of the identified positions were further confirmed by mutagenesis in three members of the subtype (UHRF1-PHD, KDM5B-PHD and KAT6A-PHD) that included pairs sharing even less than 40% sequence identity with each other. Despite low sequence similarity, mutations cause similar consequences in histone H3 binding suggesting a strong similarity in the binding mechanism, and thus justifying the subtype classification. Advisors/Committee Members: Suvobrata Chakravarty.

Subjects/Keywords: Bromodomain adjacent to Zn Finger 2A (BAZ2A); Isothermal Titration Colorimeter (ITC); Mutagenesis; PHD finger subtype; Protein expression; Protein purification; Biochemistry

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Boamah, D. (2017). Dissecting the Histone-binding Mechanism of a PHD Finger Subtype. (Doctoral Dissertation). South Dakota State University. Retrieved from https://openprairie.sdstate.edu/etd/1673

Chicago Manual of Style (16^th Edition):

Boamah, Daniel. “Dissecting the Histone-binding Mechanism of a PHD Finger Subtype.” 2017. Doctoral Dissertation, South Dakota State University. Accessed February 28, 2021. https://openprairie.sdstate.edu/etd/1673.

MLA Handbook (7^th Edition):

Boamah, Daniel. “Dissecting the Histone-binding Mechanism of a PHD Finger Subtype.” 2017. Web. 28 Feb 2021.

Vancouver:

Boamah D. Dissecting the Histone-binding Mechanism of a PHD Finger Subtype. [Internet] [Doctoral dissertation]. South Dakota State University; 2017. [cited 2021 Feb 28]. Available from: https://openprairie.sdstate.edu/etd/1673.

Council of Science Editors:

Boamah D. Dissecting the Histone-binding Mechanism of a PHD Finger Subtype. [Doctoral Dissertation]. South Dakota State University; 2017. Available from: https://openprairie.sdstate.edu/etd/1673

Open Access Theses and Dissertations