title: Hydrogen Sulfide Modulates the S-Nitrosoproteome and the Mitochondrial Morphology in Endothelial Cells.
abstract: BACKGROUND
  Hydrogen sulfide (H2S) is one of the endogenous gaseous molecules promoting the production of nitric oxide (NO) which has cardioprotective functions. However, the role of the H2S-mediated protein S-nitrosoproteome and its subsequent physiological response remains unclear.
  
  
  METHODS
  Endothelial cells EAhy 926 were treated with 50 μM of H2S for 2 hours. The NO bound S-nitrosoproteins were purified by a biotin-switch and then digested by trypsin. Resulting peptides from control and H2S treatment were separately labeled by isobaric tag for relative and absolute quantitation 114/115, quantified by liquid chromatography tandem-mass spectrometry and analyzed by ingenuity pathway analysis (IPA) software. The microP software was applied to analyze the morphological changes of mitochondria.
  
  
  RESULTS
  With the treatment of H2S, 416 S-nitrosylated proteins were identified. IPA analysis showed that these proteins were involved in five signaling pathways. The NO-bound cysteine residues and the S-nitrosylation levels (115/114) were shown for ten S-nitrosoproteins. Western blot further verified the S-nitrosylation of thioredoxin-dependant peroxide reductase, cytochrome c oxidase and cytochrome b-c1 complex that are involved in the mitochondrial signaling pathway. H2O2-induced mitochondrial swelling can be reduced by the pretreatment of H2S.
  
  
  CONCLUSIONS
  The H2S-mediated endothelial S-nitrosoproteome has been confirmed. In the present study, we have proposed the cardioprotective role of H2S via maintaining mitochondrial homeostasis.
authors.affiliation: Division of Cardiology, Ten Chan General Hospital, Chung-Li, Taoyuan.
authors.forename: Tsan-Wan
authors.initials: TW
authors.lastname: Chiu
authors.affiliation: Department of Medicine, MacKay Medical College, New Taipei City; ; Department of Anesthesiology, MacKay Memorial Hospital; ; MacKay Junior College of Medicine, Nursing and Management, Taipei.
authors.forename: Ying-Lun
authors.initials: YL
authors.lastname: Chen
authors.affiliation: Department of Pediatrics, E-Da Hospital; ; School of Medicine, College of Medicine, I-Shou University.
authors.forename: Chien-Yi
authors.initials: CY
authors.lastname: Wu
authors.affiliation: Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung.
authors.forename: Pei-Ling
authors.initials: PL
authors.lastname: Yu
authors.affiliation: Division of Family Medicine, Wan Fang Medical Center, Taipei Medical University, Taipei.
authors.forename: Ying-Hua
authors.initials: YH
authors.lastname: Shieh
authors.affiliation: Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung; ; Center for Biomarkers and Biotech Drugs; ; Center for Infectious Disease and Cancer Research, Kaohsiung Medical University; ; Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung, Taiwan.
authors.forename: Bin
authors.initials: B
authors.lastname: Huang
keywords: Endothelial cell; Hydrogen sulfide; MicroP; Mitochondria; Mitophagy; S-nitrosoproteome