Activity-based protein profiling: The serine hydrolases

• DOI: 10.1073/pnas.96.26.14694
• Publication Date: 2002-07-26T14:41:48Z
• Authors: Yongsheng Liu; Matthew P. Patric...; Benjamin F. Cravatt
• Keywords: Serine hydrolase

With the postgenome era rapidly approaching, new strategies for functional analysis of proteins are needed. To date, proteomics efforts have primarily been confined to recording variations in protein level rather than activity. The ability profile classes on basis changes their activity would greatly accelerate both assignment function and identification potential pharmaceutical targets. Here, we describe chemical synthesis utility an active-site directed probe visualizing dynamics expression entire enzyme family, serine hydrolases. By reacting this probe, a biotinylated fluorophosphonate referred as FP-biotin, with crude tissue extracts, quickly high sensitivity detect numerous hydrolases, many which display tissue-restricted patterns expression. Additionally, show that FP-biotin labels these activity-dependent manner can be followed kinetically, offering powerful means monitor simultaneously