Characterization of the sex-dependent myocardial S -nitrosothiol proteome

• Published in: American journal of physiology. Heart and circulatory physiology Vol. 310; no. 4; pp. H505 - H515
• Main Authors: Shao, Qin, Fallica, Jonathan, Casin, Kevin M., Murphy, Elizabeth, Steenbergen, Charles, Kohr, Mark J.
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 15.02.2016
• Subjects: Animals; Coronary Circulation - drug effects; Cyclophilins - metabolism; Endoplasmic Reticulum - metabolism; Female; Heart - drug effects; Male; Menopause; Mice; Mice, Inbred C57BL; Myocardial Reperfusion Injury - metabolism; Myocardium - metabolism; Nitric Oxide - biosynthesis; Nitric Oxide Synthase Type III - biosynthesis; Peptidyl-Prolyl Isomerase F; Phosphorylation; Physiology; Proteins; Proteome - drug effects; Proteomics; S-Nitrosothiols - metabolism; Sex Characteristics; Signaling and Stress Response; Womens health; ischemia-reperfusion injury; S-nitrosoglutathione reductase; cardioprotection; endothelial nitric oxide synthase; sex differences; S-nitrosylation
• ISSN: 0363-6135; 1522-1539
• DOI: 10.1152/ajpheart.00681.2015

Premenopausal women exhibit endogenous cardioprotective signaling mechanisms that are thought to result from the beneficial effects of estrogen, which we have shown to increase protein S-nitrosylation in the heart. S-nitrosylation is a labile protein modification that increases with a number of different forms of cardioprotection, including ischemic preconditioning. Herein, we sought to identify a potential role for protein S-nitrosylation in sex-dependent cardioprotection. We utilized a Langendorff-perfused mouse heart model of ischemia-reperfusion injury with male and female hearts, and S-nitrosylation-resin-assisted capture with liquid chromatography tandem mass spectrometry to identify S-nitrosylated proteins and modification sites. Consistent with previous studies, female hearts exhibited resilience to injury with a significant increase in functional recovery compared with male hearts. In a separate set of hearts, we identified a total of 177 S-nitrosylated proteins in female hearts at baseline compared with 109 S-nitrosylated proteins in male hearts. Unique S-nitrosylated proteins in the female group included the F 1 F O -ATPase and cyclophilin D. We also utilized label-free peptide analysis to quantify levels of common S-nitrosylated identifications and noted that the S-nitrosylation of sarcoplasmic/endoplasmic reticulum Ca 2+ -ATPase 2a was nearly 70% lower in male hearts compared with female, with no difference in expression. Furthermore, we found a significant increase in endothelial nitric oxide synthase expression, phosphorylation, and total nitric oxide production in female hearts compared with males, likely accounting for the enhanced S-nitrosylation protein levels in female hearts. In conclusion, we identified a number of novel S-nitrosylated proteins in female hearts that are likely to contribute to sex-dependent cardioprotection.