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

MT Forrester, JW Thompson, MW Foster… - Nature …, 2009 - nature.com
Nature biotechnology, 2009nature.com
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 …
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.
nature.com