A novel approach to identify proteins modified by nitric oxide: the HIS-TAG switch method

J Proteome Res. 2007 Aug;6(8):3224-31. doi: 10.1021/pr0701456. Epub 2007 Jul 13.

Abstract

S-nitrosylation is emerging as an important signaling mechanism that regulates a broad range of cellular functions. The recognition of Cysteine residues that undergo S-nitrosylation is crucial to elucidate how this modification modulates protein activity. We report here a novel strategy, defined His-tag switch, which allows the purification and identification of S-nitrosylated proteins and the unambiguous localization of the modified cysteine residues by mass spectrometry analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cerebellum / metabolism*
  • Female
  • Male
  • Molecular Sequence Data
  • Nerve Tissue Proteins / analysis*
  • Nerve Tissue Proteins / chemistry
  • Nitric Oxide / chemistry
  • Nitric Oxide / metabolism*
  • Nitroso Compounds / chemistry*
  • Ovalbumin / analysis
  • Ovalbumin / chemistry
  • Rats
  • Rats, Sprague-Dawley
  • S-Nitrosoglutathione / chemistry*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods

Substances

  • Nerve Tissue Proteins
  • Nitroso Compounds
  • Nitric Oxide
  • S-Nitrosoglutathione
  • Ovalbumin