subject:(Legumain)

University of Oslo

1. Nguyen, Nhan Trong. Effekten av Lipopolysakkarid (LPS) på cysteinproteaser i RAW 264,7-celler og THP-1 celler.

Degree: 2006, University of Oslo

URL: http://urn.nb.no/URN:NBN:no-17226 ; https://www.duo.uio.no/handle/10852/12130

► Ekspresjon og aktivitet av legumain ble studert i RAW 264,7-celler (en makrofaglignende leukemisk cellelinje fra mus) og THP-1 celler (en human monocyttlignende akutt leukemisk cellelinje).… (more)

▼ Ekspresjon og aktivitet av legumain ble studert i RAW 264,7-celler (en makrofaglignende leukemisk cellelinje fra mus) og THP-1 celler (en human monocyttlignende akutt leukemisk cellelinje). LPS (endotoksin) er et lipopolysakkarid som er en bestanddel av de gramnegative bakterienes cellevegg. Den toksiske delen av LPS er lipid A, som kan stimulere celler i immunsystemet. For å studere hvordan RAW 264,7-celler og THP-1 celler påvirkes av LPS, er det interessant å se på innhold av proteolytiske enzymer (cysteinproteaser legumain og cathepsin B) i endosom/lysosom. Legumain og CB er vist å bidra til antigenpresentasjon av MHC klasse II-molekyler. Ved reseptorstimulering via mikroorganismer eller cytokiner, induseres makrofager til å uttrykke MHC-II slik at de blir antigenpresenterende celler (APC). RAW 264,7-celler og THP-1 celler ble inkubert med og uten LPS i 0-72 timer. Enzymaktivitet av legumain og CB ble målt i cellelysat ved hjelp av spesifikke peptidsubstrater og beregnet i forhold til total proteininnhold. Innhold og størrelse av legumain i RAW 264,7-celler og THP-1 celler ble analysert ved immunodeteksjon (Western blot) med legumainspesifikt antistoff. Ved hjelp av Real Time-PCR analyse ble legumain m-RNA undersøkt i RAW 264,7-celler og THP-1 celler, og om legumain reguleres ved stimulering med LPS. Resultatene viste at legumain og CB enzymaktivitet er mye høyere i RAW 264,7-celler enn i THP-1 celler (for ustimulerte og stimulerte celler). Funnene tyder ikke på at LPS har noe virkning på legumain og CB enzymaktivitet, da det var ingen forskjell mellom ustimulerte og stimulerte celler. Innhold og størrelse av legumain i RAW 264,7-celler viste et proteinbånd på 37 kDa. Dette båndet kan representere en aktiv form av legumain, siden vi målte en økning i spesifikk enzymaktivitet ved samme behandling og tidspunkt. Men legumain i THP-1 celler viste et protein bånd på 50 kDa. Dette båndet kan representere en inaktiv form av legumain, siden vi så ikke bånd i området 32-37 kDa, der vi forventet å finne den aktive formen av legumain. Dette stemte med aktivitetsmålinger. Det er uklart om LPS har noe effekt på legumain m-RNA, og om man kan forvente at ekspresjon av legumain i kontroll av antigen prosessering vil bli redusert i en tilstand av endotoksintoleranse i monocytter. Advisors/Committee Members: Professor Harald T. Johansen og professor Rigmor Solberg.

Subjects/Keywords: legumain; VDP::568

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

APA (6^th Edition):

Nguyen, N. T. (2006). Effekten av Lipopolysakkarid (LPS) på cysteinproteaser i RAW 264,7-celler og THP-1 celler. (Thesis). University of Oslo. Retrieved from http://urn.nb.no/URN:NBN:no-17226 ; https://www.duo.uio.no/handle/10852/12130

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):

Nguyen, Nhan Trong. “Effekten av Lipopolysakkarid (LPS) på cysteinproteaser i RAW 264,7-celler og THP-1 celler.” 2006. Thesis, University of Oslo. Accessed March 07, 2021. http://urn.nb.no/URN:NBN:no-17226 ; https://www.duo.uio.no/handle/10852/12130.

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

MLA Handbook (7^th Edition):

Nguyen, Nhan Trong. “Effekten av Lipopolysakkarid (LPS) på cysteinproteaser i RAW 264,7-celler og THP-1 celler.” 2006. Web. 07 Mar 2021.

Vancouver:

Nguyen NT. Effekten av Lipopolysakkarid (LPS) på cysteinproteaser i RAW 264,7-celler og THP-1 celler. [Internet] [Thesis]. University of Oslo; 2006. [cited 2021 Mar 07]. Available from: http://urn.nb.no/URN:NBN:no-17226 ; https://www.duo.uio.no/handle/10852/12130.

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

Council of Science Editors:

Nguyen NT. Effekten av Lipopolysakkarid (LPS) på cysteinproteaser i RAW 264,7-celler og THP-1 celler. [Thesis]. University of Oslo; 2006. Available from: http://urn.nb.no/URN:NBN:no-17226 ; https://www.duo.uio.no/handle/10852/12130

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

University of Lund

2. Wallin, Hanna. Cellular Uptake of Cystatin C. Subcellular localisation and intracellular effects of a secreted cysteine protease inhibitor.

Degree: 2013, University of Lund

URL: https://lup.lub.lu.se/record/3991196 ; https://portal.research.lu.se/ws/files/3387001/3992325.pdf

► Cystatin C is a cysteine protease inhibitor, aimed for secretion, as it is produced with a signal peptide. Its target enzymes are thought to be… (more)

Subjects/Keywords: Pharmacology and Toxicology; Medicinal Chemistry; cathepsin; cell line; co-localisation; cystatin C; internalisation; legumain

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

APA (6^th Edition):

Wallin, H. (2013). Cellular Uptake of Cystatin C. Subcellular localisation and intracellular effects of a secreted cysteine protease inhibitor. (Doctoral Dissertation). University of Lund. Retrieved from https://lup.lub.lu.se/record/3991196 ; https://portal.research.lu.se/ws/files/3387001/3992325.pdf

Chicago Manual of Style (16^th Edition):

Wallin, Hanna. “Cellular Uptake of Cystatin C. Subcellular localisation and intracellular effects of a secreted cysteine protease inhibitor.” 2013. Doctoral Dissertation, University of Lund. Accessed March 07, 2021. https://lup.lub.lu.se/record/3991196 ; https://portal.research.lu.se/ws/files/3387001/3992325.pdf.

MLA Handbook (7^th Edition):

Wallin, Hanna. “Cellular Uptake of Cystatin C. Subcellular localisation and intracellular effects of a secreted cysteine protease inhibitor.” 2013. Web. 07 Mar 2021.

Vancouver:

Wallin H. Cellular Uptake of Cystatin C. Subcellular localisation and intracellular effects of a secreted cysteine protease inhibitor. [Internet] [Doctoral dissertation]. University of Lund; 2013. [cited 2021 Mar 07]. Available from: https://lup.lub.lu.se/record/3991196 ; https://portal.research.lu.se/ws/files/3387001/3992325.pdf.

Council of Science Editors:

Wallin H. Cellular Uptake of Cystatin C. Subcellular localisation and intracellular effects of a secreted cysteine protease inhibitor. [Doctoral Dissertation]. University of Lund; 2013. Available from: https://lup.lub.lu.se/record/3991196 ; https://portal.research.lu.se/ws/files/3387001/3992325.pdf

3. YIN JING. Novel mechanisms of programmed cell death in the protozoan parasite blastocystis.

Degree: 2009, National University of Singapore

URL: http://scholarbank.nus.edu.sg/handle/10635/16829

Subjects/Keywords: programmed cell death; protozoan; Blastocystis; proteomics; legumain; cysteine proteases

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

APA (6^th Edition):

JING, Y. (2009). Novel mechanisms of programmed cell death in the protozoan parasite blastocystis. (Thesis). National University of Singapore. Retrieved from http://scholarbank.nus.edu.sg/handle/10635/16829

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):

JING, YIN. “Novel mechanisms of programmed cell death in the protozoan parasite blastocystis.” 2009. Thesis, National University of Singapore. Accessed March 07, 2021. http://scholarbank.nus.edu.sg/handle/10635/16829.

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

MLA Handbook (7^th Edition):

JING, YIN. “Novel mechanisms of programmed cell death in the protozoan parasite blastocystis.” 2009. Web. 07 Mar 2021.

Vancouver:

JING Y. Novel mechanisms of programmed cell death in the protozoan parasite blastocystis. [Internet] [Thesis]. National University of Singapore; 2009. [cited 2021 Mar 07]. Available from: http://scholarbank.nus.edu.sg/handle/10635/16829.

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

Council of Science Editors:

JING Y. Novel mechanisms of programmed cell death in the protozoan parasite blastocystis. [Thesis]. National University of Singapore; 2009. Available from: http://scholarbank.nus.edu.sg/handle/10635/16829

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

University of Oslo

4. Ali, Zana Abdullrahman. Regulation of legumain activity by cystatin M in various cell lines.

Degree: 2008, University of Oslo

URL: http://urn.nb.no/URN:NBN:no-21470 ; https://www.duo.uio.no/handle/10852/12218 ; Fulltext https://www.duo.uio.no/bitstream/handle/10852/12218/2/Ali_masteroppgave.pdf

► Abstract HEK 293 cells were transfected with LGMN-plasmid encoding rat legumain, to establish a cell line transiently and/or stably expressing legumain. In addition, HEK 293… (more)

▼ Abstract HEK 293 cells were transfected with LGMN-plasmid encoding rat legumain, to establish a cell line transiently and/or stably expressing legumain. In addition, HEK 293 cells and a number of melanoma cell lines (from Rikshospitalet University Hospital HF) were successfully transfected with CST6-plasmid encoding human cystatin M, a potent secreted endogenous inhibitor of legumain and other cysteine proteases. Successful CST6-transfection was evaluated as increase in total inhibitory activity (IU/ml) against papain as the target enzyme in media from transfected cells versus control cells (using empty vector; pTracer). The increase in total inhibitory activity was much higher in media from the CST6-transfected HEK 293 cells than the CST6-transfected melanoma cells. In addition, legumain activity was measured in cell lysates from CST6-transfected cells, showing decreased activity compared to control cells. The legumain activity was nearly abolished in cell lysates from CST6-transfected HEK 293 cells and moderately decreased (10-41 %) in CST6-transfected melanoma cells. Interestingly, decrease in legumain activity was strongly correlated to the potency of total inhibitory activity in media from the same cells. Despite many adjustments made to optimize transfection efficacy of HEK 293 cells with the LGMN-plasmid, legumain could not be over-expressed. Cystatin M-conditioned media from CST6-transfected cells were used to treat living HEK 293, THP-1 and PC12 cells to investigate how this affected legumain activity in the cells. Legumain activity was only moderately decreased in cell lysates of HEK 293 cells treated with cystatin M-conditioned compared to control cell media. However, no regulation of legumain activity was observed in either THP-1 or PC12 cells after treatment with cystatin M-conditioned media. Cell lysates from HEK 293, THP-1 and PC12 cells were subjected to size exclusion chromatography in an attempt to characterize the molecular weight of active legumain in these cells. Surprisingly, different molecular weights were found to be responsible for legumain activity in the different cell lines. Addition of cystatin M-conditioned media to HEK 293, THP-1 and PC12 cell fractions of highest legumain activity resulted in inhibition of legumain activity. Legumain activity in HEK 293 and PC12 cells was almost completely suppressed, but surprisingly only partially inhibited in THP-1 cells. Adding of the cathepsin-inhibitors E64 and CA074 to the highest legumain activity fractions from the same cell lines resulted in no inhibition of legumain in HEK 293 and PC12 cell lysate fractions. On the contrary, the legumain activity was greatly decreased in the THP-1 cell fraction by both E64 and CA074. This is the first observation of legumain inhibition by E64 and CA074 in any cell type. This might indicate an uncharacterized form of legumain or a legumain-like enzyme in monocytes/macrophages. Advisors/Committee Members: Professor Rigmor Solberg and Professor Harald Thidemann Johansen.

Subjects/Keywords: cystein proteaser protease hemmere kreft HEK 293 celler legumain; VDP::568

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

APA (6^th Edition):

Ali, Z. A. (2008). Regulation of legumain activity by cystatin M in various cell lines. (Thesis). University of Oslo. Retrieved from http://urn.nb.no/URN:NBN:no-21470 ; https://www.duo.uio.no/handle/10852/12218 ; Fulltext https://www.duo.uio.no/bitstream/handle/10852/12218/2/Ali_masteroppgave.pdf

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):

Ali, Zana Abdullrahman. “Regulation of legumain activity by cystatin M in various cell lines.” 2008. Thesis, University of Oslo. Accessed March 07, 2021. http://urn.nb.no/URN:NBN:no-21470 ; https://www.duo.uio.no/handle/10852/12218 ; Fulltext https://www.duo.uio.no/bitstream/handle/10852/12218/2/Ali_masteroppgave.pdf.

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

MLA Handbook (7^th Edition):

Ali, Zana Abdullrahman. “Regulation of legumain activity by cystatin M in various cell lines.” 2008. Web. 07 Mar 2021.

Vancouver:

Ali ZA. Regulation of legumain activity by cystatin M in various cell lines. [Internet] [Thesis]. University of Oslo; 2008. [cited 2021 Mar 07]. Available from: http://urn.nb.no/URN:NBN:no-21470 ; https://www.duo.uio.no/handle/10852/12218 ; Fulltext https://www.duo.uio.no/bitstream/handle/10852/12218/2/Ali_masteroppgave.pdf.

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

Council of Science Editors:

Ali ZA. Regulation of legumain activity by cystatin M in various cell lines. [Thesis]. University of Oslo; 2008. Available from: http://urn.nb.no/URN:NBN:no-21470 ; https://www.duo.uio.no/handle/10852/12218 ; Fulltext https://www.duo.uio.no/bitstream/handle/10852/12218/2/Ali_masteroppgave.pdf

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

5. Ovat, Asli. Design, synthesis and evaluation of cysteine protease inhibitors.

Degree: PhD, Chemistry and Biochemistry, 2009, Georgia Tech

URL: http://hdl.handle.net/1853/33822

► Cysteine proteases are important drug targets due to their involvement in many biological processes such as protein turnover, digestion, blood coagulation, apoptosis, cell differentiation, cell… (more)

▼ Cysteine proteases are important drug targets due to their involvement in many biological processes such as protein turnover, digestion, blood coagulation, apoptosis, cell differentiation, cell signaling, and the immune response. In this thesis, we have reported the design, synthesis and evaluation of clan CA and clan CD cysteine protease inhibitors. Aza-peptidyl Michael acceptor and epoxide inhibitors for asparaginyl endopeptidases (legumains) from the bloodfluke, Schistosoma mansoni (SmAE) and the hard tick, Ixodes ricinus (IrAE) were designed and synthesized. SARs were similar, but with some notable exceptions. Both enzymes prefer disubstituted amides to monosubstituted amides in the P1' position and potency increased as we increased the hydrophobicity of the inhibitor in this position. Extending the inhibitor to P5 resulted in increased inhibitory potency, especially against IrAE, and both enzymes prefer small over large hydrophobic residues in the P2 position. Aza-peptide Michael acceptor inhibitors are more potent than aza-peptide epoxide inhibitors and, for some of these compounds, second order inhibition rate constants are the fastest yet discovered. We have also synthesized aza-peptidyl Michael acceptor and epoxide inhibitors for the parasitic cysteine proteases; cruzain, rhodesain. We have found that monosubstituted amides were favored over disubstituted amides indicating the involvement of the amide hydrogen in a H-bond network. We have shown that aza-peptide epoxides were as potent as Michael acceptors and we have obtained compounds with IC50 values as low as 20 nM. We have worked on the synthesis of heterocyclic peptidyl α-ketoamides, peptidyl ketones and aza-peptidyl ketones as calpain inhibitors. We have synthesized peptidyl α-ketoamides with nucleotide bases in the primed region to create compounds that can cross the blood-brain barrier. We have improved the potency by introducing a hydrophobic group on the adenine ring. We have obtained compounds with Ki values in the nanomolar range. We have designed peptidyl aminoketones as a new class of inhibitors for calpain. Peptidyl aminoketones were less potent than peptidyl α-ketoamides but still reasonable inhibitors of calpain that have the potential to cross the BBB. Advisors/Committee Members: Powers, James C. (Committee Chair), Christoph J. Fahrni (Committee Member), Jonathan D. Glass (Committee Member), Nicholas V. Hud (Committee Member), Sheldon W. May (Committee Member).

Subjects/Keywords: Legumain; Calpain; Protease; Cysteine; Cysteine proteinases; Cysteine proteinases Inhibitors; Protease inhibitors; Biosynthesis; Epoxy compounds

…caspases, Arg for clostripain, Asn for legumain family, Arg or Lys for gingipain family, Arg for… …proteolytic degradation of hemoglobin.29 Among these are a legumain (SmAE) that, in vitro… …ricinus33 and Haemaphysalis longicornis.42 Plant legumain functions as a processing enzyme of… …storage proteins during seed germination.43 Mammalian legumain is involved in the inhibition of… …osteoclast formation and bone resorption.44 It has also been shown that human legumain 13 is…

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

APA (6^th Edition):

Ovat, A. (2009). Design, synthesis and evaluation of cysteine protease inhibitors. (Doctoral Dissertation). Georgia Tech. Retrieved from http://hdl.handle.net/1853/33822

Chicago Manual of Style (16^th Edition):

Ovat, Asli. “Design, synthesis and evaluation of cysteine protease inhibitors.” 2009. Doctoral Dissertation, Georgia Tech. Accessed March 07, 2021. http://hdl.handle.net/1853/33822.

MLA Handbook (7^th Edition):

Ovat, Asli. “Design, synthesis and evaluation of cysteine protease inhibitors.” 2009. Web. 07 Mar 2021.

Vancouver:

Ovat A. Design, synthesis and evaluation of cysteine protease inhibitors. [Internet] [Doctoral dissertation]. Georgia Tech; 2009. [cited 2021 Mar 07]. Available from: http://hdl.handle.net/1853/33822.

Council of Science Editors:

Ovat A. Design, synthesis and evaluation of cysteine protease inhibitors. [Doctoral Dissertation]. Georgia Tech; 2009. Available from: http://hdl.handle.net/1853/33822

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