Characterization of binding properties of monoglyceride lipase inhibitors by a versatile fluorescence-based technique.
Monoglyceride lipase (MGL) is a serine hydrolase that terminates the signaling of the primary endocannabinoid, 2-arachidonoyl glycerol (2-AG). Versatile high-throughput screening methods allowing the testing of MGL inhibitors are rare, thereby limiting the development and analysis of novel inhibitors. Here we describe an improved fluorescence-based technique that is capable of determining time- and dose-dependent inhibition of MGL with one or multiple binding sites and, at the same time, is capable of revealing the reversibility of inhibitor binding in a simple kinetic assay format. Known reference compounds as well as novel inhibitors, such as JZL184 and CAY10499, were evaluated for their MGL-binding properties and potency.
Savinainen
Juha R
JR
Institute of Biomedicine/Physiology, University of Kuopio, 70211 Kuopio, Finland. juha.savinainen@uef.fi
Yoshino
Megumi
M
Minkkilä
Anna
A
Nevalainen
Tapio
T
Laitinen
Jarmo T
JT
D001665:Binding Sites; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000737:chemistry*; D057166:High-Throughput Screening Assays; D007700:Kinetics; D008994:Monoacylglycerol Lipases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011485:Protein Binding
D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D008994:Monoacylglycerol Lipases