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

• Published in: Nature biotechnology Vol. 27; no. 6; pp. 557 - 559
• Main Authors: Stamler, Jonathan S, Forrester, Michael T, Thompson, J Will, Foster, Matthew W, Nogueira, Leonardo, Moseley, M Arthur
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.06.2009; Nature Publishing Group
• Subjects: Agriculture; Analytical, structural and metabolic biochemistry; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Biotin; Biotin - metabolism; brief-communication; Cell Line; Cellular biology; Cysteine - analogs & derivatives; Cysteine - metabolism; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Genetic aspects; Glyceraldehyde-3-Phosphate Dehydrogenases - metabolism; Humans; Life Sciences; Mass Spectrometry; Nitric oxide; Physiological aspects; Proteins; Proteome - analysis; Proteomics; Proteomics - methods; S-Nitrosothiols - metabolism; Scientific method; Sensitivity and Specificity; Sulfhydryl Compounds - metabolism; United States; Nitrosylation; Analysis method; Protein; Proteomics
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt.1545

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.