Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

CiteULike is a free service for managing and discovering scholarly references - click here to get started.

Sign In to gain access to subscriptions and/or personal tools.
Journal of Biomolecular Screening
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
1087057108319738v1
13/6/468    most recent
Right arrow References
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via Google Scholar
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Keller, P. M.
Right arrow Articles by Schwartz, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Keller, P. M.
Right arrow Articles by Schwartz, B.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

A High-Throughput Screen for Endothelial Lipase Using HDL as Substrate

Paul M. Keller

GSK Screening & Compound Profiling, Collegeville, Pennsylvania

Timothy Rust

GSK Screening & Compound Profiling, Collegeville, Pennsylvania

Dennis J. Murphy

GSK Cardiovascular Biochemistry, Upper Merion, Pennsylvania

Rosalie Matico

Biological Reagents & Assay Development, Collegeville, Pennsylvania

John J. Trill

Biological Reagents & Assay Development, Collegeville, Pennsylvania

John A. Krawiec

GSK Cardiovascular Biology, Upper Merion, Pennsylvania

Anthony Jurewicz

GSK Screening & Compound Profiling, Collegeville, Pennsylvania

Michael Jaye

GSK Cardiovascular Biology, Upper Merion, Pennsylvania

Mark Harpel

GSK Cardiovascular Biochemistry, Upper Merion, Pennsylvania

Sara Thrall

Biological Reagents & Assay Development, Collegeville, Pennsylvania

Benjamin Schwartz

Biological Reagents & Assay Development, Collegeville, Pennsylvania, benjamin.2.schwartz{at}gsk.com

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-µL, 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. (Journal of Biomolecular Screening 2008:468-475)

Key Words: endothelial lipase • HDL • interfacial enzymes • coupled enzyme assay • alkylation protection

This version was published on July 1, 2008

Journal of Biomolecular Screening, Vol. 13, No. 6, 468-475 (2008)
DOI: 10.1177/1087057108319738
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?