title: Resorufin butyrate as a soluble and monomeric high-throughput substrate for a triglyceride lipase.
abstract: Triglyceride lipases such as lipoprotein lipase, endothelial lipase, and hepatic lipase play key roles in controlling the levels of plasma lipoprotein. Accordingly, small-molecule modulation of these species could alter patient lipid profiles with corresponding health effects. Screening of these enzymes for small-molecule therapeutics has historically involved the use of lipid-based particles to mimic native substrates. However, particle-based artifacts can complicate the discovery of therapeutic molecules. As a simplifying solution, the authors sought to develop an approach involving a soluble and monomeric lipase substrate. Using purified bovine lipoprotein lipase as a model system, they show that the hydrolysis of resorufin butyrate can be fluorescently monitored to give a robust assay (Z' > 0.8). Critically, using parallel approaches, they show that resorufin butyrate is soluble and monomeric under assay conditions. The presented assay should be useful as a simple and inexpensive primary or secondary screen for the discovery of therapeutic lipase modulators.
authors.affiliation: Department of In Vitro Sciences, Merck Frosst Centre for Therapeutic Research, Kirkland, Quebec, Canada.
authors.forename: Vincent
authors.initials: V
authors.lastname: Lam
authors.forename: Martin
authors.initials: M
authors.lastname: Henault
authors.forename: Karine
authors.initials: K
authors.lastname: Khougaz
authors.forename: Louis-Jacques
authors.initials: LJ
authors.lastname: Fortin
authors.forename: Marc
authors.initials: M
authors.lastname: Ouellet
authors.forename: Roman
authors.initials: R
authors.lastname: Melnyk
authors.forename: Anthony
authors.initials: A
authors.lastname: Partridge
mesh_terms: D000818:Animals; D002087:Butyrates / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D057166:High-Throughput Screening Assays / Q000379:methods*; D008071:Lipoprotein Lipase / Q000032:analysis* / Q000737:chemistry; D010078:Oxazines / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002087:Butyrates; D010078:Oxazines; C014180:resorufin; D008071:Lipoprotein Lipase