Proteomic analysis of S-nitrosylation and denitrosylation by resin-assisted capture

Nat Biotechnol. 2009 Jun;27(6):557-9. doi: 10.1038/nbt.1545. Epub 2009 May 31.


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.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Biotin / metabolism
  • Cell Line
  • Cysteine / analogs & derivatives
  • Cysteine / metabolism
  • Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism
  • Humans
  • Mass Spectrometry
  • Proteome / analysis*
  • Proteomics / methods*
  • S-Nitrosothiols / metabolism*
  • Sensitivity and Specificity
  • Sulfhydryl Compounds / metabolism*


  • Proteome
  • S-Nitrosothiols
  • Sulfhydryl Compounds
  • Biotin
  • S-nitrosocysteine
  • Glyceraldehyde-3-Phosphate Dehydrogenases
  • Cysteine