Targeted Protein Capture for Analysis of Electrophile-Protein Adducts

摘要: Proteomic analyses of protein-electrophile adducts generally employ affinity capture the adduct moiety, which enables global analyses, but is poorly suited to targeted studies specific proteins. We describe a molecular-probe approach study modifications molecular chaperone heat-shock protein 90 (Hsp90), regulates diverse client Noncovalent with biotinyl analog HSP90 inhibitor geldanamycin detection native isoforms Hsp90α and Hsp90β their phosphorylated forms. applied this probe map quantify formed on Hsp90 by 4-hydroxynonenal (HNE) in RKO cells. This was also measure kinetics site-specific adduction selected residues. A protein-selective broadly applicable for analysis electrophile biological effects.