Expression of the human soluble epoxide hydrolase in Escherichia coli by auto-induction for the study of high-throughput inhibition assays
| Autor: | Seung-Jin Ma, In-Hae Kim, Kosuke Nishi |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Epoxide hydrolase 2
Insecta Immunoblotting Biology medicine.disease_cause Cell Line Substrate Specificity Inhibitory Concentration 50 High-Throughput Screening Assays Escherichia coli medicine Animals Humans Enzyme Inhibitors Enzyme inducer Epoxide Hydrolases chemistry.chemical_classification Molecular biology Enzyme assay Enzyme Solubility chemistry Biochemistry Enzyme inhibitor Cell culture Enzyme Induction biology.protein Electrophoresis Polyacrylamide Gel Biotechnology |
| Zdroj: | Protein Expression and Purification. 69:34-38 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2009.09.013 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect