title: Measurement of protein S-nitrosylation during cell signaling.
abstract: S-Nitrosylation, the modification of a cysteine thiol by a nitric oxide (NO) group, has emerged as an important posttranslational modification of signaling proteins. An impediment to studying the regulation of cell signaling by S-nitrosylation has been the technical challenge of detecting endogenously S-nitrosylated proteins. Detection of S-nitrosylated proteins is difficult because the S-NO bond is labile and therefore can be lost or gained artifactually during sample preparation. Nevertheless, several methods have been developed to measure endogenous protein S-nitrosylation, including the biotin switch assay and the chemical reduction/chemiluminescence assay. This chapter describes these two methods and provides examples of how they have been used successfully to elucidate the role of protein S-nitrosylation in cell physiology and pathophysiology.
authors.affiliation: Department of Medicine and Cell Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Joan B
authors.initials: JB
authors.lastname: Mannick
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Schonhoff
mesh_terms: D000818:Animals; D002468:Cell Physiological Phenomena; D002477:Cells / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D026403:S-Nitrosothiols / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D011506:Proteins; D026403:S-Nitrosothiols