Clemente_2012_J.Biomol.Screen_17_629

Reference

Title : Screening and characterization of human monoglyceride lipase active site inhibitors using orthogonal binding and functional assays - Clemente_2012_J.Biomol.Screen_17_629
Author(s) : Clemente JC , Nulton E , Nelen M , Todd MJ , Maguire D , Schalk-Hihi C , Kuo LC , Zhang SP , Flores CM , Kranz JK
Ref : J Biomol Screen , 17 :629 , 2012
Abstract :

Endocannabinoids such as 2-arachidonylglycerol (2-AG) are ligands for cannabinoid receptors that contribute to the transmission and modulation of pain signals. The antinociceptive effect of exogenous 2-AG suggests that inhibition of monoglyceride lipase (MGLL), the enzyme responsible for degrading 2-AG and arresting signaling, may be a target for pain modulation. Here we describe the characterization of MGLL ligands following a high-throughput screening campaign. Ligands were discovered using ThermoFluor, a label-free affinity-based screening tool that measures ligand binding via modulation of protein thermal stability. A kinetic fluorescent assay using the substrate 4-methylcoumarin butyrate was used to counterscreen confirmed HTS positives. A comparison of results from binding and inhibition assays allowed elucidation of compound mechanism of action. We demonstrate the limit of each technology and the benefits of using orthogonal assay techniques in profiling compounds.

PubMedSearch : Clemente_2012_J.Biomol.Screen_17_629
PubMedID: 22496098

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Citations formats

Clemente JC, Nulton E, Nelen M, Todd MJ, Maguire D, Schalk-Hihi C, Kuo LC, Zhang SP, Flores CM, Kranz JK (2012)
Screening and characterization of human monoglyceride lipase active site inhibitors using orthogonal binding and functional assays
J Biomol Screen 17 :629

Clemente JC, Nulton E, Nelen M, Todd MJ, Maguire D, Schalk-Hihi C, Kuo LC, Zhang SP, Flores CM, Kranz JK (2012)
J Biomol Screen 17 :629