Information about most Saccharomyces cerevisiae genes, whether newly discovered or previously identified, is very limited. Currently, much emphasis has been placed on gene analysis at the level of nucleic acids such as the study of mRNA gene expression or genome variation. However, gene function is often deduced at the level of protein analysis, which in general has not been subjected to high throughput techniques. We propose to develop high throughput methods and protein chips for the large-scale biochemical analysis of protein function in yeast. A set of yeast strains will be prepared that express yeast ORFs as GST fusions under the control of a galactose-inducible promoter; we will subsequently use high throughput methods for the large-scale expression of the yeast proteins. The proteins will be adhered to novel microwell chips that we are developing and analyzed for biochemical activities. Specifically, we will use in vitro binding and/or enzymatic assays to identify targets of GTP-binding proteins, phospholipid binding protein and protein phosphatases. We will also further analyze protein kinases and their in vitro substrates using protein microchip techniques. We expect this project to have a significant impact on the scientific community. The information generated will be maintained in a database that is accessible over the Internet. Our plasmids and yeast strains will be made available to all interested researchers. Most importantly the technology developed is expected to have an enormous impact on the study of yeast and other organisms.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM062480-04S1
Application #
6880509
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Edmonds, Charles G
Project Start
2001-03-01
Project End
2005-12-31
Budget Start
2004-03-01
Budget End
2005-12-31
Support Year
4
Fiscal Year
2004
Total Cost
$122,625
Indirect Cost
Name
Yale University
Department
Physiology
Type
Schools of Arts and Sciences
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Li, Xiyan; Snyder, Michael P (2016) Can heavy isotopes increase lifespan? Studies of relative abundance in various organisms reveal chemical perspectives on aging. Bioessays 38:1093-1101
Yang, Grace Xiaolu; Li, Xiyan; Snyder, Michael (2012) Investigating metabolite-protein interactions: an overview of available techniques. Methods 57:459-66
Chen, Rui; Snyder, Michael (2012) Systems biology: personalized medicine for the future? Curr Opin Pharmacol 12:623-8
Li, Xiyan; Snyder, Michael (2011) Analyzing In Vivo Metabolite-Protein Interactions By Large-Scale Systematic Analyses. Curr Protoc Chem Biol 3:181-196
Mok, Janine; Zhu, Xiaowei; Snyder, Michael (2011) Dissecting phosphorylation networks: lessons learned from yeast. Expert Rev Proteomics 8:775-86
Li, Xiyan; Gianoulis, Tara A; Yip, Kevin Y et al. (2010) Extensive in vivo metabolite-protein interactions revealed by large-scale systematic analyses. Cell 143:639-50
Chen, Rui; Snyder, Michael (2010) Yeast proteomics and protein microarrays. J Proteomics 73:2147-57
Mok, Janine; Im, Hogune; Snyder, Michael (2009) Global identification of protein kinase substrates by protein microarray analysis. Nat Protoc 4:1820-7
Hall, David A; Ptacek, Jason; Snyder, Michael (2007) Protein microarray technology. Mech Ageing Dev 128:161-7
Gelperin, Daniel M; White, Michael A; Wilkinson, Martha L et al. (2005) Biochemical and genetic analysis of the yeast proteome with a movable ORF collection. Genes Dev 19:2816-26

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