Detection of protein S-nitrosylation with the biotin-switch technique.
Protein S-nitrosylation, the posttranslational modification of cysteine thiols to form S-nitrosothiols, is a principle mechanism of nitric oxide-based signaling. Studies have demonstrated myriad roles for S-nitrosylation in organisms from bacteria to humans, and recent efforts have greatly advanced our scientific understanding of how this redox-based modification is dynamically regulated during physiological and pathophysiological conditions. The focus of this review is the biotin-switch technique (BST), which has become a mainstay assay for detecting S-nitrosylated proteins in complex biological systems. Potential pitfalls and modern adaptations of the BST are discussed, as are future directions for this assay in the burgeoning field of protein S-nitrosylation.
Forrester
Michael T
MT
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
Foster
Matthew W
MW
Benhar
Moran
M
Stamler
Jonathan S
JS
D000818:Animals; D001710:Biotin / Q000737:chemistry* / Q000378:metabolism; D002457:Cell Extracts / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation / Q000379:methods* / Q000639:trends; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D009569:Nitric Oxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D011499:Protein Processing, Post-Translational / Q000502:physiology; D026403:S-Nitrosothiols / Q000737:chemistry*; D015398:Signal Transduction; D013438:Sulfhydryl Compounds / Q000737:chemistry
D002457:Cell Extracts; D026403:S-Nitrosothiols; D013438:Sulfhydryl Compounds; D009569:Nitric Oxide; D001710:Biotin; D003545:Cysteine