Assay and inhibition of diacylglycerol lipase activity.
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 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.
Center for Drug Discovery, Northeastern University, 360 Huntington Avenue, Boston, MA 02115, USA. meghanryan.johnston@gmail.com
Meghan
M
Johnston
Shachi R
SR
Bhatt
Surina
S
Sikka
Richard W
RW
Mercier
Jay M
JM
West
Alexandros
A
Makriyannis
S John
SJ
Gatley
Richard I
RI
Duclos
D000818:Animals; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors*; D051379:Mice; D015394:Molecular Structure; D013329:Structure-Activity Relationship
D004791:Enzyme Inhibitors; D008071:Lipoprotein Lipase