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University of Georgia

1. Wallace, Erica Renee. Using proteomics as a tool for the study of micronutrient metabolism.

Degree: 2014, University of Georgia

Proteomics is the comprehensive and systematic study of the proteome, which is defined as the set of all expr essed proteins in a cell, tissue or organism. The standard method for quantitative proteome analysis combines protein separation by two -dimensional gel electrophoresis (2DGE) with mass spectrometric (MS) identification of selected protein spots. Because these techniques allow one to study the full protein complement of a cell or tissue at the time of isolation, they are beneficial for identifying the pool of proteins whose expression is related to a particular treatment and to each other. This paper will pr esent two proteomic studies of micronutrient metabolism. In the first study, the effect of zinc deficiency on protein expression in a colon adenocarcinoma (Caco -2) cell-culture model was investigated. In the second study, proteomics techniques were used to identify alterations in cellular protein expression, due to transfection with a selenoprotein construct ( pro-fs) in Madin-Darby Canine Kidney cells.

Subjects/Keywords: Two-dimensional gel electrophoresis; Zinc; Selenium; Caco-2; Madin- Darby Canine Kidney; Zinc-deficiency; pro-fs; HIV-1; proteomics; mass spectrometry

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

APA (6th Edition):

Wallace, E. R. (2014). Using proteomics as a tool for the study of micronutrient metabolism. (Thesis). University of Georgia. Retrieved from http://hdl.handle.net/10724/22258

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

Chicago Manual of Style (16th Edition):

Wallace, Erica Renee. “Using proteomics as a tool for the study of micronutrient metabolism.” 2014. Thesis, University of Georgia. Accessed January 18, 2021. http://hdl.handle.net/10724/22258.

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

MLA Handbook (7th Edition):

Wallace, Erica Renee. “Using proteomics as a tool for the study of micronutrient metabolism.” 2014. Web. 18 Jan 2021.

Vancouver:

Wallace ER. Using proteomics as a tool for the study of micronutrient metabolism. [Internet] [Thesis]. University of Georgia; 2014. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/10724/22258.

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

Council of Science Editors:

Wallace ER. Using proteomics as a tool for the study of micronutrient metabolism. [Thesis]. University of Georgia; 2014. Available from: http://hdl.handle.net/10724/22258

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


University of Georgia

2. Su, Guoping. Demonstration of a molecular interaction between thioredoxin and the pro-fs gene product of HIV-1, a mimic of NF kappa B.

Degree: 2014, University of Georgia

A programmed –1 frameshift in the HIV-1 protease gene has been previously demonstrated. The sequence encoded in the overlapping –1 reading frame, named pro-fs, has significant similarity to the DNA binding loop of NF-êB, which is also the peptide known to bind thioredoxin (Trx) as part of the process of NF-êB activation. The hypothesis that the putative HIV-1 pro-fs gene product functions by mimicry of NF-êB via interacting with Trx was tested by coimmunoprecipitation and GST-pull down assay in vitro. Both experiments consistently showed that pro-fs binds human Trx in vitro; and the binding affinity is apparently stronger with Trx-wt than with Trx-CS, in which the two Cys residues in the active center of Trx were mutated to Ser. The fact that pro-fs interacts with Trx-CS rules out the possibility that the interaction between pro-fs and Trx is just due to cross-linkage via formation of an intermolecular disulfide bond between the Cys residues of pro-fs (the Sec residues in pro-fs sequence are mutated to Cys for bacterial expression) and the Cys residues in the active site of Trx. Due to the nature of the –1 frameshift mechanism, pro-fs and the HIV-1 protease share the first 20 amino acid residues at the N-terminus. Since both the retroviral protease and NF-êB function as dimers, we investigated whether pro-fs also dimerizes in living mammalian cells by fluorescent resonance energy transfer (FRET) analysis using confocal microscopy. Fluorescence microscopy was utilized to investigate the hypothesis that pro-fs is a nuclear protein, based upon its high pI and mimicry of NF-êB. The results demonstrated that pro-fs localizes in cell nuclei and forms oligomers. FRET analysis was also used to study the interaction between pro-fs and Trx in living cells. The results showed that phorbol 12-myristate 13-acetate (PMA) treatment of the 293T cells induces the nuclear translocation of Trx, and pro-fs binds to both Trx-wt and Trx-CS in PMA-stimulated cells. Together with the results of co-immunoprecipitation and GST-pull down assay, it suggests that Trx-pro-fs binding is a structurally specific interaction that involves multiple amino acid residues in the interactive region.

Subjects/Keywords: Frameshift; retrovirus; HIV-1; protease; pro-fs; NF kappa B; thioredoxin; PMA; FRET; confocal microscopy; cysteine; dimer; oligomer; GST-pull down; co-immunoprecipitation.

Record DetailsSimilar RecordsGoogle PlusoneFacebookTwitterCiteULikeMendeleyreddit

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

APA (6th Edition):

Su, G. (2014). Demonstration of a molecular interaction between thioredoxin and the pro-fs gene product of HIV-1, a mimic of NF kappa B. (Thesis). University of Georgia. Retrieved from http://hdl.handle.net/10724/21379

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

Chicago Manual of Style (16th Edition):

Su, Guoping. “Demonstration of a molecular interaction between thioredoxin and the pro-fs gene product of HIV-1, a mimic of NF kappa B.” 2014. Thesis, University of Georgia. Accessed January 18, 2021. http://hdl.handle.net/10724/21379.

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

MLA Handbook (7th Edition):

Su, Guoping. “Demonstration of a molecular interaction between thioredoxin and the pro-fs gene product of HIV-1, a mimic of NF kappa B.” 2014. Web. 18 Jan 2021.

Vancouver:

Su G. Demonstration of a molecular interaction between thioredoxin and the pro-fs gene product of HIV-1, a mimic of NF kappa B. [Internet] [Thesis]. University of Georgia; 2014. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/10724/21379.

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

Council of Science Editors:

Su G. Demonstration of a molecular interaction between thioredoxin and the pro-fs gene product of HIV-1, a mimic of NF kappa B. [Thesis]. University of Georgia; 2014. Available from: http://hdl.handle.net/10724/21379

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

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