title: Determination of lipoprotein lipase activity using a novel fluorescent lipase assay.
abstract: A novel, real-time, homogeneous fluorogenic lipoprotein lipase (LPL) assay was developed using a commercially available substrate, the EnzChek lipase substrate, which is solubilized in Zwittergent. The triglyceride analog substrate does not fluoresce, owing to apposition of fluorescent and fluorescent quenching groups at the sn-1 and sn-2 positions, respectively, fluorescence becoming unquenched upon release of the sn-1 BODIPY FA derivative following hydrolysis. Increase in fluorescence intensity at 37°C was proportional to LPL concentration. The assay was more sensitive than a similar assay using 1,2-O-dilauryl-rac-glycero-3-glutaric acid-(6-methylresorufin ester) and was validated in biological samples, including determination of LPL-specific activity in postheparin mouse plasma. The simplicity and reproducibility of the assay make it ideal for in vitro, high-throughput screening for inhibitors and activators of LPL, thus expediting discovery of drugs of potential clinical value.
authors.affiliation: Department of Cell Biology, State University of New York Downstate Medical Center, Brooklyn, NY 11203, USA.
authors.forename: Debapriya
authors.initials: D
authors.lastname: Basu
authors.forename: Jahan
authors.initials: J
authors.lastname: Manjur
authors.forename: Weijun
authors.initials: W
authors.lastname: Jin
mesh_terms: D000818:Animals; D002460:Cell Line; D057075:Enzyme Assays / Q000379:methods*; D006801:Humans; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D008071:Lipoprotein Lipase / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D014280:Triglycerides / Q000378:metabolism
chemical_list: D014280:Triglycerides; D008049:Lipase; D008071:Lipoprotein Lipase