| A high-throughput screen for endothelial lipase using HDL as substrate. | |
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MedLine Citation:
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PMID: 18566479 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Endothelial lipase (EL) is a 482-amino-acid protein from the triglyceride lipase gene family that uses a Ser-His-Asp triad for catalysis. Its expression in endothelial cells and preference for phospholipids rather than triglycerides are unique. Animal models in which it is overexpressed or knocked out indicate EL levels are inversely correlated with high-density lipoprotein cholesterol (HDL-C). HDL-C is commonly referred to as the good form of cholesterol because it is involved in the reverse cholesterol transport pathway, in which excess cholesterol is effluxed from peripheral tissues for excretion or reabsorption. Thus, EL inhibition in humans is expected to lead to increases in HDL levels and possibly a decrease in cardiovascular disease. To discover inhibitors of EL, a coupled assay for EL has been developed, using its native substrate, HDL. Hydrolysis of HDL by EL yields free fatty acids, which are coupled through acyl-CoA synthetase, acyl-CoA oxidase, and horseradish peroxidase to produce the fluorescent species resorufin. This assay was developed into a 5-microL, 1536-well assay format, and a high-throughput screen was executed against the GSK collection. In addition to describing the screening results, novel post-HTS mechanism-of-action studies were developed for EL and applied to 1 of the screening hits as an example. |
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Authors:
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Paul M Keller; Timothy Rust; Dennis J Murphy; Rosalie Matico; John J Trill; John A Krawiec; Anthony Jurewicz; Michael Jaye; Mark Harpel; Sara Thrall; Benjamin Schwartz |
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Publication Detail:
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Type: Journal Article Date: 2008-06-19 |
Journal Detail:
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Title: Journal of biomolecular screening Volume: 13 ISSN: 1087-0571 ISO Abbreviation: J Biomol Screen Publication Date: 2008 Jul |
Date Detail:
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Created Date: 2008-07-28 Completed Date: 2008-09-03 Revised Date: 2011-05-23 |
Medline Journal Info:
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Nlm Unique ID: 9612112 Medline TA: J Biomol Screen Country: United States |
Other Details:
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Languages: eng Pagination: 468-75 Citation Subset: IM |
Affiliation:
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GSK Screening & Compound Profiling, Collegeville, Pennsylvania, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acyl-CoA Oxidase
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metabolism Adenosine Triphosphate / metabolism Animals Biological Assay CHO Cells Coenzyme A Ligases / metabolism Cricetinae Cricetulus Fatty Acids, Nonesterified / metabolism Horseradish Peroxidase / metabolism Humans Hydrolysis Kinetics Lipase / chemistry, genetics, isolation & purification, metabolism* Lipoproteins, HDL / metabolism* Models, Biological Oxazines / metabolism Recombinant Proteins / chemistry, metabolism Substrate Specificity |
| Chemical | |
Reg. No./Substance:
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0/Fatty Acids, Nonesterified; 0/Lipoproteins, HDL; 0/Oxazines; 0/Recombinant Proteins; 56-65-5/Adenosine Triphosphate; 635-78-9/resorufin; EC 1.11.1.-/Horseradish Peroxidase; EC 1.3.3.6/Acyl-CoA Oxidase; EC 3.1.1.3/Lipase; EC 6.2.1.-/Coenzyme A Ligases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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