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Identification of potential S‐nitrosylation sites in the myocardium
Author(s) -
Kohr Mark Jeffrey,
Sun Junhui,
Aponte Angel,
Wang Guanghui,
Gucek Marjan,
Murphy Elizabeth,
Steenbergen Charles
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.1094.4
Subject(s) - s nitrosylation , cysteine , chemistry , proteome , nitric oxide , nitrosylation , biochemistry , proteomics , microbiology and biotechnology , biology , gene , enzyme , organic chemistry
S‐nitrosylation (SNO) is a reversible protein modification that results from the covalent attachment of nitric oxide to a thiol group of a cysteine residue. Numerous methodologies have been developed to detect SNO proteins, but very few SNO sites have been identified in complex samples. Therefore, the objective of this study is to identify potential SNO sites in the myocardium. We utilized SNO‐resin assisted capture (SNO‐RAC) with mass spectrometry in order to determine SNO sites. Briefly, crude mouse heart homogenate was incubated with or without the S‐nitrosylating agent GSNO (1 mM) and a modified biotin switch was performed. SNO‐RAC analysis identified 952 unique SNO proteins in GSNO‐treated homogenates, many of which contained multiple SNO sites. These proteins all show the potential for endogenous SNO formation. Of the 952 SNO proteins identified with GSNO treatment, 116 were identified to be endogenously SNO in control homogenates in the absence of GSNO. Gene ontology analysis revealed that many different pathways are targeted by SNO, including myocardial contraction, metabolism, cellular signaling and cell death. This study provides novel information regarding the cardiac SNO proteome and provides additional information on the sites of SNO for key proteins involved in many different myocardial processes. Supported by NIH grants 1F32HL096142 and 5R01HL039752, and the NIH Intramural Program.