| pubmed-article:21397586 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:21397586 | lifeskim:mentions | umls-concept:C0023764 | lld:lifeskim |
| pubmed-article:21397586 | lifeskim:mentions | umls-concept:C0242485 | lld:lifeskim |
| pubmed-article:21397586 | lifeskim:mentions | umls-concept:C0182400 | lld:lifeskim |
| pubmed-article:21397586 | pubmed:issue | 2 | lld:pubmed |
| pubmed-article:21397586 | pubmed:dateCreated | 2011-5-18 | lld:pubmed |
| pubmed-article:21397586 | pubmed:abstractText | Modulating the activity of lipases involved in the metabolism of plasma lipoproteins is an attractive approach for developing lipid raising/lowering therapies to treat cardiovascular disease. Identifying small molecule inhibitors for these membrane-active enzymes, however, is complicated by difficulties associated with measuring lipase activity and inhibition at the water-membrane interface; substrate and compound dynamics at the particle interface have the potential to confound data interpretation. Here, we describe a novel ELISA-based lipase activity assay that employs as "bait" a biotinylated active-site probe that irreversibly binds to the catalytic active-site serine of members of the triacylglycerol lipase family (hepatic lipase, lipoprotein lipase, and endothelial lipase) in solution with high affinity. Detection of "captured" (probe-enzyme) complexes on streptavidin-coated plates using labeled secondary antibodies to specific primary antibodies offers several advantages over conventional assays, including the ability to eliminate enzyme-particle and compound-particle effects; specifically measure lipase activity in complex mixtures in vitro; preferentially identify active-site-directed inhibitors; and distinguish between reversible and irreversible inhibitors through a simple assay modification. Using EL as an exemplar, we demonstrate the versatility of this assay both for high-throughput screening and for compound mechanism-of-action studies. | lld:pubmed |
| pubmed-article:21397586 | pubmed:language | eng | lld:pubmed |
| pubmed-article:21397586 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21397586 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:21397586 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21397586 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21397586 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21397586 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21397586 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21397586 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21397586 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:21397586 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:21397586 | pubmed:month | Jul | lld:pubmed |
| pubmed-article:21397586 | pubmed:issn | 1096-0309 | lld:pubmed |
| pubmed-article:21397586 | pubmed:author | pubmed-author:MelnykRoman... | lld:pubmed |
| pubmed-article:21397586 | pubmed:author | pubmed-author:PartridgeAnth... | lld:pubmed |
| pubmed-article:21397586 | pubmed:author | pubmed-author:KINGM OMO | lld:pubmed |
| pubmed-article:21397586 | pubmed:author | pubmed-author:HenaultMartin... | lld:pubmed |
| pubmed-article:21397586 | pubmed:author | pubmed-author:WangZhaoyinZ | lld:pubmed |
| pubmed-article:21397586 | pubmed:author | pubmed-author:LiLianhaiL | lld:pubmed |
| pubmed-article:21397586 | pubmed:copyrightInfo | Copyright © 2011 Elsevier Inc. All rights reserved. | lld:pubmed |
| pubmed-article:21397586 | pubmed:issnType | Electronic | lld:pubmed |
| pubmed-article:21397586 | pubmed:day | 15 | lld:pubmed |
| pubmed-article:21397586 | pubmed:volume | 414 | lld:pubmed |
| pubmed-article:21397586 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:21397586 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:21397586 | pubmed:pagination | 254-60 | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:meshHeading | pubmed-meshheading:21397586... | lld:pubmed |
| pubmed-article:21397586 | pubmed:year | 2011 | lld:pubmed |
| pubmed-article:21397586 | pubmed:articleTitle | An activity-based probe for high-throughput measurements of triacylglycerol lipases. | lld:pubmed |
| pubmed-article:21397586 | pubmed:affiliation | Department of In Vitro Sciences, Merck Frosst Centre for Therapeutic Research, 16711 Trans Canada Highway, Kirkland, Quebec, Canada. | lld:pubmed |
| pubmed-article:21397586 | pubmed:publicationType | Journal Article | lld:pubmed |
