2-Arachidonoylglycerol is a substrate for butyrylcholinesterase: A potential mechanism for extracellular endocannabinoid regulation

2-Arachidonoylglycerol (2-AG) is a component of the endocannabinoid receptor pathway and is primarily hydrolyzed by monoacylglycerol lipase (MAGL) in vivo. We found that the non-specific serine esterase, butyrylcholinesterase (BChE), can hydrolyze 2-AG with reasonable affinity and may present a new compensatory mechanism for endocannabinoid regulation. In vitro hydrolysis reactions of 2-AG with equine BChE were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) positive/negative electrospray ionization (ESI +/-) to measure the formation of arachidonic acid (AA) and the loss of 2-AG over time (min). The resulting Michaelis-Menten approximations reveal that BChE has affinity towards 2-AG in phosphate buffer at neutral pH (7.4). The calculated V-max, K-m and k(cat) were 12.1 nmol s(-1), 57.5 mu M, and 0.074 s(-1), respectively, which produced a diffusion-controlled rate of association (k(cat)/K-m) of 1.3 x 10(3) - M-1 s(-1). Human BChE 2-AG hydrolysis was measured by immunoprecipitating BChE from fresh plasma and monitoring 2-AG loss and AA formation over time. These findings show that BChE can hydrolyze 2-AG which may be evidence of a more specific role for BChE in endocannabinoid regulation. (C) 2013 Elsevier Inc. All rights reserved.