Chemoproteomic discovery of AADACL1 as a regulator of human platelet activation

Stephen P Holly; Jae Won Chang; Weiwei Li; Sherry Niessen; Ryan M Phillips; Raymond Piatt; Justin L Black; Matthew C Smith; Yacine Boulaftali; Andrew S Weyrich; Wolfgang Bergmeier; Benjamin F Cravatt; Leslie V Parise

doi:10.1016/j.chembiol.2013.07.011

Chemoproteomic discovery of AADACL1 as a regulator of human platelet activation

Chem Biol. 2013 Sep 19;20(9):1125-34. doi: 10.1016/j.chembiol.2013.07.011. Epub 2013 Aug 29.

Authors

Stephen P Holly¹, Jae Won Chang, Weiwei Li, Sherry Niessen, Ryan M Phillips, Raymond Piatt, Justin L Black, Matthew C Smith, Yacine Boulaftali, Andrew S Weyrich, Wolfgang Bergmeier, Benjamin F Cravatt, Leslie V Parise

Affiliation

¹ Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. Electronic address: sholly@med.unc.edu.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Blood Platelets / metabolism*
Carbamates / chemistry
Carbamates / metabolism
Carbamates / pharmacology
Carboxylic Ester Hydrolases / antagonists & inhibitors
Carboxylic Ester Hydrolases / genetics
Carboxylic Ester Hydrolases / metabolism*
Cell Line
Enzyme Inhibitors / chemistry
Enzyme Inhibitors / metabolism
Enzyme Inhibitors / pharmacology
Fibrinogen / metabolism
Humans
Lipid Metabolism / drug effects
Megakaryocytes / cytology
Megakaryocytes / drug effects
Megakaryocytes / metabolism
Metabolomics
Platelet Activation / drug effects
Platelet Aggregation / drug effects
Platelet Glycoprotein GPIIb-IIIa Complex / metabolism
Protein Binding
Protein Kinase C / metabolism
Proteomics
RNA Interference
RNA, Small Interfering / metabolism
Sterol Esterase
rap1 GTP-Binding Proteins / metabolism

Substances

Carbamates
Enzyme Inhibitors
Platelet Glycoprotein GPIIb-IIIa Complex
RNA, Small Interfering
Fibrinogen
Protein Kinase C
Carboxylic Ester Hydrolases
NCEH1 protein, human
Sterol Esterase
rap1 GTP-Binding Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding