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Design and Synthesis of Quenched Activity‐based Probes for Diacylglycerol Lipase and α,β‐Hydrolase Domain Containing Protein 6
Author(s) -
van Rooden E. J.,
Kohsiek M.,
Kreekel R.,
van Esbroeck A. C. M.,
van den Nieuwendijk A. M. C. H.,
Janssen A. P. A.,
van den Berg R. J. B. H. N.,
Overkleeft H. S.,
van der Stelt M.
Publication year - 2018
Publication title -
chemistry – an asian journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.18
H-Index - 106
eISSN - 1861-471X
pISSN - 1861-4728
DOI - 10.1002/asia.201800452
Subject(s) - diacylglycerol lipase , diacylglycerol kinase , monoacylglycerol lipase , fluorophore , lipase , chemistry , hydrolase , biochemistry , serine hydrolase , endocannabinoid system , fluorescence , serine , circular dichroism , moiety , biophysics , enzyme , stereochemistry , biology , protein kinase c , receptor , physics , quantum mechanics
Diacylglycerol lipases (DAGL) are responsible for the biosynthesis of the endocannabinoid 2‐arachidonoylglycerol. The fluorescent activity‐based probes DH379 and HT‐01 have been previously shown to label DAGLs and to cross‐react with the serine hydrolase ABHD6. Here, we report the synthesis and characterization of two new quenched activity‐based probes 1 and 2 , the design of which was based on the structures of DH379 and HT‐01, respectively. Probe 1 contains a BODIPY‐FL and a 2,4‐dinitroaniline moiety as a fluorophore–quencher pair, whereas probe 2 employs a Cy5‐fluorophore and a cAB40‐quencher. The fluorescence of both probes was quenched with relative quantum yields of 0.34 and 0.0081, respectively. The probes showed target inhibition as characterized in activity‐based protein profiling assays using human cell‐ and mouse brain lysates, but were unfortunately not active in living cells, presumably due to limited cell permeability.