The rapid development of proteomic techniques in the post-genomics era has allowed
much attention to be paid to the understanding of molecular mechanisms of functional regulatory
proteins. This amazingly fast transformation from know-how to know-why has allowed the
determination of if functional regulation is based on up- or down-regulation, or based on any
particular chemical modification on the amino acid residues of protein targets.
Here, different biologically important regulatory proteomes or protein targets were
investigated using comprehensive combinations of proteomics techniques. A large emphasis was
placed on expression techniques suitable for protein and protein binding partner affinity
purification for targeted analysis of phospho-proteins. The main protein of interest in these
particular studies was an important transcriptional factor in cardiac development, serum response
factor-1. Several proteomics methods were examined and, along with the difficulties
encountered, the results are presented. Small molecule-protein interaction was examined with the
cell cycle regulatory protein Cdc25B incubated with an inhibitor. Post-translational
modification-related functional proteomics were also studied, first in a rodent early hemorrhage
model wherein S-nitrosylation of plasma proteins were determined, then the phosphorylation of
serum response factor-1. Finally, top-down functional proteome mapping was represented by a
detailed and very successful analysis of the proteins found enriched in brain tumor cell
pseudopodia.
This dissertation illustrates the vast range of potential of proteomics technologies in
studying function-related biological systems. Meanwhile, some pioneering work using targeted
functional proteomics strategies was achieved, and the results will help in part to advance the
field of targeted functional proteomics, the combination of cell and molecular biological
techniques with chemical, affinity and mass spectrometric approaches to study regulation of
biological systems.
| Identifer | oai:union.ndltd.org:PITT/oai:PITTETD:etd-06162007-121003 |
| Date | 22 June 2007 |
| Creators | Zhang, Zhe |
| Contributors | Yu Jiang, Ph.D., Michael A Zemaitis, Ph.D., Robert B Gibbs, Ph.D., Jorge L Sepulveda M.D., Ph.D., Billy W Day, Ph.D. |
| Publisher | University of Pittsburgh |
| Source Sets | University of Pittsburgh |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.pitt.edu/ETD/available/etd-06162007-121003/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Pittsburgh or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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