A novel assay for monoacylglycerol hydrolysis suitable for high-throughput screening.
A simple assay for monoacylglycerol hydrolysis suitable for high-throughput screening is described. The assay uses [(3)H]2-oleoylglycerol as substrate, with the tritium label in the glycerol part of the molecule and the use of phenyl sepharose gel to separate the hydrolyzed product ([(3)H]glycerol) from substrate. Using cytosolic fractions derived from rat cerebella as a source of hydrolytic activity, the assay gives the appropriate pH profile and sensitivity to inhibition with compounds known to inhibit hydrolysis of this substrate. The assay could also be adapted to a 96-well plate format, using C6 cells as the source of hydrolytic activity. Thus the assay is simple and appropriate for high-throughput screening of inhibitors of monoacylglycerol hydrolysis.
Brengdahl
Johan
J
Department of Pharmacology and Clinical Neuroscience, Umeå University, SE-901 87 Umeå, Sweden.
Fowler
Christopher J
CJ
D000818:Animals; D001331:Automation; D001671:Biochemistry / Q000379:methods; D001681:Biological Assay / Q000379:methods*; D019411:Clinical Laboratory Techniques; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006868:Hydrolysis; D007700:Kinetics; D008994:Monoacylglycerol Lipases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D050178:Monoglycerides / Q000378:metabolism*; D051381:Rats; D012685:Sepharose / Q000737:chemistry; D014316:Tritium / Q000378:metabolism
D004791:Enzyme Inhibitors; D050178:Monoglycerides; D014316:Tritium; D012685:Sepharose; D008994:Monoacylglycerol Lipases