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

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

Author information

1
Department of Biochemistry, Duke University Medical Center, Durham, North Carolina, USA.

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.

PMID:
19483679
PMCID:
PMC2891235
DOI:
10.1038/nbt.1545
[Indexed for MEDLINE]
Free PMC Article

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