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.
|Canterbury, Jesse D; Merrihew, Gennifer E; MacCoss, Michael J et al. (2014) Comparison of data acquisition strategies on quadrupole ion trap instrumentation for shotgun proteomics. J Am Soc Mass Spectrom 25:2048-59|
|Bateman, Nicholas W; Goulding, Scott P; Shulman, Nicholas J et al. (2014) Maximizing peptide identification events in proteomic workflows using data-dependent acquisition (DDA). Mol Cell Proteomics 13:329-38|
|Hsieh, Edward J; Bereman, Michael S; Durand, Stanley et al. (2013) Effects of column and gradient lengths on peak capacity and peptide identification in nanoflow LC-MS/MS of complex proteomic samples. J Am Soc Mass Spectrom 24:148-53|
|Glukhova, Veronika A; Tomazela, Daniela M; Findlay, Geoffrey D et al. (2013) Rapid assessment of RNAi-mediated protein depletion by selected reaction monitoring mass spectrometry. J Proteome Res 12:3246-54|
|Bereman, Michael S; Canterbury, Jesse D; Egertson, Jarrett D et al. (2012) Evaluation of front-end higher energy collision-induced dissociation on a benchtop dual-pressure linear ion trap mass spectrometer for shotgun proteomics. Anal Chem 84:1533-9|
|Bereman, Michael S; Tomazela, Daniela M; Heins, Hillary S et al. (2012) A method to determine the kinetics of multiple proteins in human infants with respiratory distress syndrome. Anal Bioanal Chem 403:2397-402|
|Gilligan, Diana M; Finney, Greg L; Rynes, Eric et al. (2011) Comparative proteomics reveals deficiency of NHE-1 (Slc9a1) in RBCs from the beta-adducin knockout mouse model of hemolytic anemia. Blood Cells Mol Dis 47:85-94|
|Bereman, Michael S; Egertson, Jarrett D; MacCoss, Michael J (2011) Comparison between procedures using SDS for shotgun proteomic analyses of complex samples. Proteomics 11:2931-5|
|Hsieh, Edward J; Hoopmann, Michael R; MacLean, Brendan et al. (2010) Comparison of database search strategies for high precursor mass accuracy MS/MS data. J Proteome Res 9:1138-43|
|Tomazela, Daniela M; Patterson, Bruce W; Hanson, Elizabeth et al. (2010) Measurement of human surfactant protein-B turnover in vivo from tracheal aspirates using targeted proteomics. Anal Chem 82:2561-7|
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