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Assay and inhibition of diacylglycerol lipase activity.

ABSTRACT: A series of N-formyl-?-amino acid esters of ?-lactone derivatives structurally related to tetrahydrolipstatin (THL) and O-3841 were synthesized that inhibit human and murine diacylglycerol lipase (DAGL) activities. New ether lipid reporter compounds were developed for an in vitro assay to efficiently screen inhibitors of 1,2-diacyl-sn-glycerol hydrolysis and related lipase activities using fluorescence resonance energy transfer (FRET). A standardized thin layer chromatography (TLC) radioassay of diacylglycerol lipase activity utilizing the labeled endogenous substrate [1?-(14)C]1-stearoyl-2-arachidonoyl-sn-glycerol with phosphorimaging detection was used to quantify inhibition by following formation of the initial product [1?-(14)C]2-arachidonoylglycerol and further hydrolysis under the assay conditions to [1-(14)C]arachidonic acid.

SUBMITTER: Johnston M

PROVIDER: S-EPMC3389199 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature

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Assay and inhibition of diacylglycerol lipase activity.

Johnston Meghan M Bhatt Shachi R SR Sikka Surina S Mercier Richard W RW West Jay M JM Makriyannis Alexandros A Gatley S John SJ Duclos Richard I RI

Bioorganic & medicinal chemistry letters 20120606 14

A series of N-formyl-α-amino acid esters of β-lactone derivatives structurally related to tetrahydrolipstatin (THL) and O-3841 were synthesized that inhibit human and murine diacylglycerol lipase (DAGL) activities. New ether lipid reporter compounds were developed for an in vitro assay to efficiently screen inhibitors of 1,2-diacyl-sn-glycerol hydrolysis and related lipase activities using fluorescence resonance energy transfer (FRET). A standardized thin layer chromatography (TLC) radioassay of ...[more]

PMID: 22738638

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