title: Characterization of the sex-dependent myocardial S-nitrosothiol proteome.
abstract: 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 F1FO-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.
authors.affiliation: Department of Cardiology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China; Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, Maryland;
authors.forename: Qin
authors.initials: Q
authors.lastname: Shao
authors.affiliation: Department of Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland; and.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Fallica
authors.affiliation: Department of Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland; and.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Casin
authors.affiliation: Systems Biology Center, National Heart, Lung, and Blood Institute, Bethesda, Maryland.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Murphy
authors.affiliation: Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, Maryland;
authors.forename: Charles
authors.initials: C
authors.lastname: Steenbergen
authors.affiliation: Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, Maryland; Department of Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland; and mkohr1@jhu.edu.
authors.forename: Mark J
authors.identifier: http://orcid.org/0000-0002-6034-5962
authors.initials: MJ
authors.lastname: Kohr
mesh_terms: D000818:Animals; D003326:Coronary Circulation / Q000187:drug effects*; D000081406:Peptidyl-Prolyl Isomerase F; D021983:Cyclophilins / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005260:Female; D006321:Heart / Q000187:drug effects*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015428:Myocardial Reperfusion Injury / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis; D010766:Phosphorylation; D020543:Proteome / Q000187:drug effects*; D026403:S-Nitrosothiols / Q000378:metabolism*; D012727:Sex Characteristics
chemical_list: D000081406:Peptidyl-Prolyl Isomerase F; C000631446:PPIF protein, mouse; D020543:Proteome; D026403:S-Nitrosothiols; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; D021983:Cyclophilins
keywords: S-nitrosoglutathione reductase; S-nitrosylation; cardioprotection; endothelial nitric oxide synthase; ischemia-reperfusion injury; sex differences