The proteomic core will provide cutting edge discovery protein biochemistry and proteomics tools to address key mechanistic questions, in support ofthe underlying concept that disease-induced changes in autophagy lead to alterations in protein quantity, abundance of specific isoforms and post-translational modifications, which drive cellular phenotype. The core will provide discovery based approaches for protein quantification and the identification of post-translational modifications including phosphorylation, 0-GlcNAcylation and cysteine oxidative modifications for the detailed analysis of a number of subproteomes and protein complexes. The core will employ its recently modified method that allows the site specific quantification and comparison of S-nitrosation, S-glutathionylation and sulfonic acid, the three reversible cysteine modifications. As well, the core will provide multiplex assays for the absolute quantification of key proteins involved in autophagy, lysosomal function, mitochondrial biogenesis and nutrient sensing using multiple reaction monitoring, a targeted mass spectrometry based method. The panels developed will quantify the total protein concentration as well as differentiate between the various protein isoforms which arise from different genes or splice variants of a single gene and for a subset, specific regulatory posttranslational modification.

Public Health Relevance

The purpose of the Protein Chemistry and Proteomic Core (Core B) is to apply cutting edge discovery protein biochemistry and proteomics tools to address key mechanistic questions from each PPG project. Core B will provide crucial experimental services in support of the underlying concept that disease-induced changes in protein quantify and PTMs dictate the underlying autophagy status, its regulation by different signaling events and ultimately myocyte function. Being able to therapeutically manipulate autophagy will allow us to address protection and heart failure.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
7P01HL112730-02
Application #
8683230
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
Project End
Budget Start
2014-07-21
Budget End
2015-06-30
Support Year
2
Fiscal Year
2014
Total Cost
$262,313
Indirect Cost
$108,011
Name
Cedars-Sinai Medical Center
Department
Type
DUNS #
075307785
City
Los Angeles
State
CA
Country
United States
Zip Code
90048
Gottlieb, Roberta A; Bernstein, Daniel (2016) Mitochondrial remodeling: Rearranging, recycling, and reprogramming. Cell Calcium 60:88-101
Stotland, Aleksandr; Gottlieb, Roberta A (2016) α-MHC MitoTimer mouse: In vivo mitochondrial turnover model reveals remarkable mitochondrial heterogeneity in the heart. J Mol Cell Cardiol 90:53-8
Gottlieb, Roberta A; Pourpirali, Somayeh (2016) Lost in translation: miRNAs and mRNAs in ischemic preconditioning and ischemia/reperfusion injury. J Mol Cell Cardiol 95:70-7
Lam, Maggie P Y; Venkatraman, Vidya; Xing, Yi et al. (2016) Data-Driven Approach To Determine Popular Proteins for Targeted Proteomics Translation of Six Organ Systems. J Proteome Res 15:4126-4134
Holewinski, Ronald J; Parker, Sarah J; Matlock, Andrea D et al. (2016) Methods for SWATHâ„¢: Data Independent Acquisition on TripleTOF Mass Spectrometers. Methods Mol Biol 1410:265-79
Sin, Jon; Andres, Allen M; Taylor, David J R et al. (2016) Mitophagy is required for mitochondrial biogenesis and myogenic differentiation of C2C12 myoblasts. Autophagy 12:369-80
Gottlieb, Roberta A; Andres, Allen M; Sin, Jon et al. (2015) Untangling autophagy measurements: all fluxed up. Circ Res 116:504-14
Chung, Heaseung Sophia; Murray, Christopher I; Venkatraman, Vidya et al. (2015) Dual Labeling Biotin Switch Assay to Reduce Bias Derived From Different Cysteine Subpopulations: A Method to Maximize S-Nitrosylation Detection. Circ Res 117:846-57
Gurney, Michael A; Huang, Chengqun; Ramil, Jennifer M et al. (2015) Measuring cardiac autophagic flux in vitro and in vivo. Methods Mol Biol 1219:187-97
Delbridge, Lea M D; Mellor, Kimberley M; Taylor, David J R et al. (2015) Myocardial autophagic energy stress responses--macroautophagy, mitophagy, and glycophagy. Am J Physiol Heart Circ Physiol 308:H1194-204

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