title: Chemoproteomic discovery of AADACL1 as a regulator of human platelet activation.
abstract: A comprehensive knowledge of the platelet proteome is necessary for understanding thrombosis and for envisioning antiplatelet therapies. To discover other biochemical pathways in human platelets, we screened platelets with a carbamate library designed to interrogate the serine hydrolase subproteome and used competitive activity-based protein profiling to map the targets of active carbamates. We identified an inhibitor that targets arylacetamide deacetylase-like 1 (AADACL1), a lipid deacetylase originally identified in invasive cancers. Using this compound, along with highly selective second-generation inhibitors of AADACL1, metabolomics, and RNA interference, we show that AADACL1 regulates platelet aggregation, thrombus growth, RAP1 and PKC activation, lipid metabolism, and fibrinogen binding to platelets and megakaryocytes. These data provide evidence that AADACL1 regulates platelet and megakaryocyte activation and highlight the value of this chemoproteomic strategy for target discovery in platelets.
authors.affiliation: Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. Electronic address: sholly@med.unc.edu.
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Holly
authors.forename: Jae Won
authors.initials: JW
authors.lastname: Chang
authors.forename: Weiwei
authors.initials: W
authors.lastname: Li
authors.forename: Sherry
authors.initials: S
authors.lastname: Niessen
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Phillips
authors.forename: Raymond
authors.initials: R
authors.lastname: Piatt
authors.forename: Justin L
authors.initials: JL
authors.lastname: Black
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Smith
authors.forename: Yacine
authors.initials: Y
authors.lastname: Boulaftali
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Weyrich
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Bergmeier
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
authors.forename: Leslie V
authors.initials: LV
authors.lastname: Parise
mesh_terms: D001792:Blood Platelets / Q000378:metabolism*; D002219:Carbamates / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects; D008533:Megakaryocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D055432:Metabolomics; D015539:Platelet Activation / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism; D040901:Proteomics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D002787:Sterol Esterase; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism
chemical_list: D002219:Carbamates; D004791:Enzyme Inhibitors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D034741:RNA, Small Interfering; D005340:Fibrinogen; D011493:Protein Kinase C; D002265:Carboxylic Ester Hydrolases; C515677:NCEH1 protein, human; D002787:Sterol Esterase; D020827:rap1 GTP-Binding Proteins