S-Nitrosylation Proteome Profile of Peripheral Blood Mononuclear Cells in Human Heart Failure

SPRATT, HEIDI M.; GUPTA, SHIVALI; KUYUMCU-MARTINEZ, MUGE N.; GARG, NISHA JAIN; KOO, SUE-JIE; STAFFORD, SUSAN; ZAGO, MARIA P.; WIKTOROWICZ, JOHN E.; SOMAN, KIZHAKE V.; PETERSEN, JOHN R.; BRASIER, ALLAN R.

International Journal of Proteomics

Hindawi Publishing Corporation

Lugar: El Cairo; Año: 2016 vol. 2016 p. 1 - 19

2090-2166

Nitric oxide (NO) protects the heart against ischemic injury; however, NO- and superoxide-dependent S-nitrosylation (S-NO) of cysteines can affect function of target proteins and play a role in disease outcome. We employed 2D-GE with thiol-labeling FL-maleimide dye and MALDI-TOF MS/MS to capture the quantitative changes in abundance and S-NO proteome of HF patients (versus healthy controls, n = 30/group). We identified 93 differentially abundant (59-increased/34-decreased) and 111 S-NO-modified (63-increased/48-decreased) protein spots, respectively, in HF subjects (versus controls, fold-change | ≥1.5|, p ≤ 0.05). Ingenuity pathway analysis of proteome datasets suggested that the pathways involved in phagocytes´ migration, free radical production, and cell death were activated and fatty acid metabolism was decreased in HF subjects. Multivariate adaptive regression splines modeling of datasets identified a panel of proteins that will provide >90% prediction success in classifying HF subjects. Proteomic profiling identified ATP-synthase, thrombospondin-1 (THBS1), and vinculin (VCL) as top differentially abundant and S-NO-modified proteins, and these proteins were verified by Western blotting and ELISA in different set of HF subjects. We conclude that differential abundance and S-NO modification of proteins serve as a mechanism in regulating cell viability and free radical production, and THBS1 and VCL evaluation will potentially be useful in the prediction of heart failure.