The insulin signaling pathway is essential for the regulation of glucose homeostasis, contributes to the regulation of energy substrate metabolism, and is intimately linked to aging and longevity. Because of the prevalence of obesity and type-2 diabetes within industrialized societies, understanding the molecular mechanism of this pathway is an increasing biomedical priority. Insulin like molecules alter system-wide protein turnover, yet the effects on individual proteins is unclear. A change in a protein's turnover can alter the stoichiometry of the active protein and ultimately affect the efficiency of a pathway. Because of the potential functional consequence of protein turnover and the importance of insulin signaling, we propose to study both these processes in the nematode Caenorhabditis elegans using quantitative proteornics.
Specific Aim 1 is to develop quantitative stable isotope tracer methodology for both the directed and undirected analysis of protein turnover in C. elegans.
Specific Aim 2 will measure the turnover of proteins involved in the insulin/insulin growth factor-1 (IGF-1) signaling pathway of C. elegans in response to the inhibition and overexpression of ligands of the insulin receptor (DAF-2).
Specific Aim 3 is to measure the effect of insulin like molecules on the tissue-specific intracellular metabolism of the nematode C. elegans using stable isotope labeling and isotopomer distribution analysis. The successfulcompletion of this proposal will produce methodological frameworkfor studying protein turnover in C. elegans and provide foundation for extending these technologies to higher organisms. Furthermore, the proteome-wide measurement of protein turnover in response to insulin-like molecules will produce an experimental dataset for probing the underlying mechanism of insulin signaling.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK069386-07
Application #
8241898
Study Section
Enabling Bioanalytical and Biophysical Technologies Study Section (EBT)
Program Officer
Sechi, Salvatore
Project Start
2006-04-15
Project End
2013-11-30
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
7
Fiscal Year
2012
Total Cost
$302,268
Indirect Cost
$106,248
Name
University of Washington
Department
Genetics
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
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