biostudies-literatureXu HNIGMS NIH HHS89-96https://www.ebi.ac.uk/biostudies/studies/S-EPMC3566859biostudies-literatureUnknown9The fluorophosphonate (FP) moiety attached to a biotin tag is a prototype chemical probe used to quantitatively analyze and enrich active serine hydrolases in complex proteomes in an approach called activity-based protein profiling (ABPP). In this study we have designed a novel synthetic route to a known FP probe linked by polyethylene glycol to a biotin tag (FP-PEG-biotin). Our route markedly increases the efficiency of the probe synthesis and overcomes several problems of a prior synthesis. As a proof of principle, FP-PEG-biotin was evaluated against isolated protein mixtures and different rat-tissue homogenates, showing its ability to specifically target serine hydrolases. We also assessed the ability of FP-PEG-biotin to compete with substrates that have high enzyme turnover rates. The Beilstein journal of organic chemistryAn improved synthesis of a fluorophosphonate-polyethylene glycol-biotin probe and its use against competitive substrates.PMC3566859R01 GM037188Sabit HShowalter HDHXu HAmidon GLfalseAn improved synthesis of a fluorophosphonate-polyethylene glycol-biotin probe and its use against competitive substrates.The fluorophosphonate (FP) moiety attached to a biotin tag is a prototype chemical probe used to quantitatively analyze and enrich active serine hydrolases in complex proteomes in an approach called activity-based protein profiling (ABPP). In this study we have designed a novel synthetic route to a known FP probe linked by polyethylene glycol to a biotin tag (FP-PEG-biotin). Our route markedly increases the efficiency of the probe synthesis and overcomes several problems of a prior synthesis. As a proof of principle, FP-PEG-biotin was evaluated against isolated protein mixtures and different rat-tissue homogenates, showing its ability to specifically target serine hydrolases. We also assessed the ability of FP-PEG-biotin to compete with substrates that have high enzyme turnover rates. The 2013-01-01T00:00:00Z20132025-04-04T10:12:00.386Z2019-03-27T01:04:29ZS-EPMC35668592340070010.3762/bjoc.9.12