Discovering new pathways in the heart during hypoxia: Proteomic approaches to identify the cardiac prolyl hydroxalome and to analyze protein stability

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Authors

• A Stoehr
• AM Evangelista
• Y Yang
• S Patel
• A Aponte
• G Wang
• J Boylston
• Y Lin
• M Gucek
• J Zhou
• E Murphy

Abstract

Hypoxia activates signaling pathways that allow the organism to adapt to reduced oxygen levels. This research aims to understand hypoxic mechanisms in cardiac diseases which might be useful to develop therapies. Prolyl hydroxylases (PHD) are oxygen-dependent enzymes which add OH groups to prolines on HIF-1α that targets it for degradation. The goal of this study was to (I) identify new targets of PHDs in the heart, (II) test whether inhibition of prolyl hydroxylation alters protein stability and (III) investigate biological consequences. Proteomics (Orbitrap LC-MS/MS) was used to identify 18 unique prolyl hydroxylated proteins in rat hearts. To address whether prolyl hydroxylation influences protein stability and to transfer the results to human, we used a cell culture model of iPS cell-derived cardiomyocytes and Stable Isotopic Labeling with Amino Acids (SILAC). In the presence of the PHD inhibitor dimethyloxalylglycine (DMOG, 1 mM) we identified 97 proteins with differences in light peptides and 53 proteins with differences in heavy peptides between DMOG and vehicle using mass spectroscopy (FDR=0.1; p

Scientific Focus Area: Systems Biology

This page was last updated on Friday, March 26, 2021