title: Proteomic analysis of S-nitrosylation and denitrosylation by resin-assisted capture.
abstract: We have modified the biotin switch assay for protein S-nitrosothiols (SNOs), using resin-assisted capture (SNO-RAC). Compared with existing methodologies, SNO-RAC requires fewer steps, detects high-mass S-nitrosylated proteins more efficiently, and facilitates identification and quantification of S-nitrosylated sites by mass spectrometry. When combined with iTRAQ labeling, SNO-RAC revealed that intracellular proteins may undergo rapid denitrosylation on a global scale. This methodology is readily adapted to analyzing diverse cysteine-based protein modifications, including S-acylation.
authors.affiliation: Department of Biochemistry, Duke University Medical Center, Durham, North Carolina, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Forrester
authors.forename: J Will
authors.initials: JW
authors.lastname: Thompson
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Foster
authors.forename: Leonardo
authors.initials: L
authors.lastname: Nogueira
authors.forename: M Arthur
authors.initials: MA
authors.lastname: Moseley
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Stamler
mesh_terms: D001710:Biotin / Q000378:metabolism; D002460:Cell Line; D003545:Cysteine / Q000031:analogs & derivatives / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D026403:S-Nitrosothiols / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013438:Sulfhydryl Compounds / Q000378:metabolism*
chemical_list: D020543:Proteome; D026403:S-Nitrosothiols; D013438:Sulfhydryl Compounds; D001710:Biotin; C021315:S-nitrosocysteine; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D003545:Cysteine