title: Superfamily-wide portrait of serine hydrolase inhibition achieved by library-versus-library screening.
abstract: Serine hydrolases (SHs) are one of the largest and most diverse enzyme classes in mammals. They play fundamental roles in virtually all physiological processes and are targeted by drugs to treat diseases such as diabetes, obesity, and neurodegenerative disorders. Despite this, we lack biological understanding for most of the 110+ predicted mammalian metabolic SHs, in large part because of a dearth of assays to assess their biochemical activities and a lack of selective inhibitors to probe their function in living systems. We show here that the vast majority (> 80%) of mammalian metabolic SHs can be labeled in proteomes by a single, active site-directed fluorophosphonate probe. We exploit this universal activity-based assay in a library-versus-library format to screen 70+ SHs against 140+ structurally diverse carbamates. Lead inhibitors were discovered for ∼40% of the screened enzymes, including many poorly characterized SHs. Global profiles identified carbamate inhibitors that discriminate among highly sequence-related SHs and, conversely, enzymes that share inhibitor sensitivity profiles despite lacking sequence homology. These findings indicate that sequence relatedness is not a strong predictor of shared pharmacology within the SH superfamily. Finally, we show that lead carbamate inhibitors can be optimized into pharmacological probes that inactivate individual SHs with high specificity in vivo.
authors.affiliation: The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Bachovchin
authors.forename: Tianyang
authors.initials: T
authors.lastname: Ji
authors.forename: Weiwei
authors.initials: W
authors.lastname: Li
authors.forename: Gabriel M
authors.initials: GM
authors.lastname: Simon
authors.forename: Jacqueline L
authors.initials: JL
authors.lastname: Blankman
authors.forename: Alexander
authors.initials: A
authors.lastname: Adibekian
authors.forename: Heather
authors.initials: H
authors.lastname: Hoover
authors.forename: Sherry
authors.initials: S
authors.lastname: Niessen
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
mesh_terms: D002219:Carbamates / Q000494:pharmacology* / Q000627:therapeutic use; D020134:Catalytic Domain; D055808:Drug Discovery / Q000379:methods*; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005459:Fluorides; D006801:Humans; D006867:Hydrolases / Q000037:antagonists & inhibitors*; D015335:Molecular Probes; D010710:Phosphates; D011485:Protein Binding; D020543:Proteome; D012694:Serine; D054852:Small Molecule Libraries / Q000494:pharmacology*; D013379:Substrate Specificity
chemical_list: D002219:Carbamates; D004791:Enzyme Inhibitors; D015335:Molecular Probes; D010710:Phosphates; D020543:Proteome; D054852:Small Molecule Libraries; C012980:fluorophosphate; D012694:Serine; D006867:Hydrolases; D005459:Fluorides