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


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.

SUBMITTER: Forrester MT 

PROVIDER: S-EPMC2891235 | biostudies-literature | 2009 Jun

REPOSITORIES: biostudies-literature

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

Forrester Michael T MT   Thompson J Will JW   Foster Matthew W MW   Nogueira Leonardo L   Moseley M Arthur MA   Stamler Jonathan S JS  

Nature biotechnology 20090531 6


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 readil  ...[more]

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