Detection and proteomic identification of S-nitrosated proteins in human hepatocytes.
The S-nitrosation of protein thiols is a redox-based posttranslational modification that modulates protein function and cell phenotype. Although the detection of S-nitrosated proteins is problematical because of the lability of S-nitrosothiols, an increasing range of proteins has been shown to undergo S-nitrosation with the improvement of molecular tools. This chapter describes the methodology used to identify potential targets of S-nitrosation in cultured primary human hepatocytes using proteomic approaches. This methodology is based on the biotin switch method, which labels S-nitrosated proteins with an affinity tag, allowing their selective detection and proteomic identification.
López-Sánchez
Laura M
LM
Liver Research Unit, Hospital Universitario Reina Sofía, Córdoba, Spain.
Corrales
Fernando J
FJ
De La Mata
Manuel
M
Muntané
Jordi
J
Rodríguez-Ariza
Antonio
A
D002478:Cells, Cultured; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D015538:Nitrosation; D011506:Proteins / Q000032:analysis* / Q000145:classification / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D026403:S-Nitrosothiols / Q000032:analysis* / Q000145:classification / Q000378:metabolism*
D011506:Proteins; D026403:S-Nitrosothiols