| Expression of the human soluble epoxide hydrolase in Escherichia coli by auto-induction for the study of high-throughput inhibition assays. | |
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MedLine Citation:
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PMID: 19782755 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Soluble epoxide hydrolase (sEH) is a key enzyme involved in the metabolism of epoxy fatty acid mediators such as epoxyeicosatrienoic acids with emerging roles in the regulations of hypertension and inflammation. Inhibitors of human sEH (hsEH) are effective drug candidates for the treatment of cardiovascular diseases. Preparation of hsEH for enzyme inhibition studies has been carried out by using baculovirus expression system. We herein explored the feasibility of expression of hsEH in Escherichia coli cells for the study of high-throughput screening assays of enzyme inhibitors, because the bacterial expression system is easier to handle and more cost-effective than the baculovirus expression system. The functional target enzyme was successfully produced in prokaryotic expression system by an auto-induction method and exhibited comparable enzyme activity to that yielded in baculovirus expression system. The bacterial-hsEH showed similar sensitivity to the baculovirus-hsEH against six reported inhibitors. Overalls indicate that bacterial expression of hsEH employed in the present study is useful for preparing enzymatically active hsEH, leading to effective performance of high-throughput screening assay of hsEH inhibitors and to rapid identification of novel drug candidates for the treatment of cardiovascular diseases. |
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Authors:
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Kosuke Nishi; In-Hae Kim; Seung-Jin Ma |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2009-09-25 |
Journal Detail:
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Title: Protein expression and purification Volume: 69 ISSN: 1096-0279 ISO Abbreviation: Protein Expr. Purif. Publication Date: 2010 Jan |
Date Detail:
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Created Date: 2009-11-06 Completed Date: 2010-01-26 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9101496 Medline TA: Protein Expr Purif Country: United States |
Other Details:
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Languages: eng Pagination: 34-8 Citation Subset: IM |
Affiliation:
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Industry and Academic Cooperation Laboratory for Drug Development, Mokpo National University, Muan, Jeonnam 534-729, South Korea. kosuke.nishi@tecan.com |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line Electrophoresis, Polyacrylamide Gel Enzyme Induction / drug effects Enzyme Inhibitors / chemistry, pharmacology Epoxide Hydrolases / antagonists & inhibitors, biosynthesis*, isolation & purification Escherichia coli / cytology, drug effects, metabolism* High-Throughput Screening Assays / methods* Humans Immunoblotting Inhibitory Concentration 50 Insects Solubility / drug effects Substrate Specificity / drug effects |
| Chemical | |
Reg. No./Substance:
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0/Enzyme Inhibitors; EC 3.3.2.-/Epoxide Hydrolases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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