Investigation of the role of cytochrome P4502B4 active site residues in substrate metabolism based on crystal structures of the ligand-bound enzyme
文献类型：期刊论文
| 作者 | Hernandez, Cynthia E.; Kumar, Santosh; Liu, Hong ; Halpert, James R.
|
| 刊名 | ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
 |
| 出版日期 | 2006-11-01 |
| 卷号 | 455期号:1页码:61-67 |
| 关键词 | cytochrome P450 structure function relationships site-directed mutagenesis enzyme catalysis and inhibition P450 2134 crystal structure |
| ISSN号 | 0003-9861 |
| DOI | 10.1016/j.abb.2006.08.024 |
| 文献子类 | Article |
| 英文摘要 | Based on the X-ray crystal structures of 4-(4-chlorophenyl)imidazole (4-CPI)- and bifonazole (BIF)-bound P450 2134, eight active site mutants at six positions were created in an N-terminal modified construct termed 2B4dH and characterized for enzyme inhibition and catalysis. I363A showed a > 4-fold decrease in differential inhibition by BIT and 4-CPI (IC50,BIF/IC50,4-CPI). F296A, T302A, I363A, V367A, and V477A showed a >= 2-fold decreased k(cat) for 7-ethoxy-4-trifluoromethylcoumarin O-deethylation, whereas V367A and V477F showed an altered K-m. T302A, V367L, and V477A showed > 4-fold decrease in total testosterone hydroxylation, whereas I363A, V367A, and V477F showed altered sterco- and regioselectivity. Interestingly, I363A showed a >= 150-fold enhanced k(cat)/K-m with testosterone, and yielded a new metabolite. Furthermore, testosterone docking into three-dimensional models of selected mutants based on the 4-CPI-bound structure suggested a re-positioning of residues 363 and 477 to yield products. In conclusion, our results suggest that the 4-CPI-bound 2B4dH/H226Y crystal structure is an appropriate model for predicting enzyme catalysis. Published by Elsevier Inc. |
| WOS关键词 | DIRECTED MUTAGENESIS ; AMINO-ACID ; RECOGNITION SITES ; SPECIFICITY ; BINDING ; INHIBITION ; EXPRESSION ; RESOLUTION ; IMIDAZOLE ; INSIGHT |
| 资助项目 | NIEHS NIH HHS[ES 03619] ; NIEHS NIH HHS[R01 ES003619] ; NIEHS NIH HHS[ES 06676] ; NIEHS NIH HHS[P30 ES006676] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biophysics |
| 语种 | 英语 |
| WOS记录号 | WOS:000241768100007 |
| 出版者 | ELSEVIER SCIENCE INC |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/273451]  |
| 专题 | 中国科学院上海药物研究所 |
| 通讯作者 | Kumar, Santosh |
| 作者单位 | 1.Univ Texas, Med Branch, Dept Pharmacol & Toxicol, Galveston, TX 77555 USA 2.Chinese Acad Sci, Shanghai Inst Mat Med, Drug Discovery & Design Ctr, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Hernandez, Cynthia E.,Kumar, Santosh,Liu, Hong,et al. Investigation of the role of cytochrome P4502B4 active site residues in substrate metabolism based on crystal structures of the ligand-bound enzyme[J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS,2006,455(1):61-67. |
| APA | Hernandez, Cynthia E.,Kumar, Santosh,Liu, Hong,&Halpert, James R..(2006).Investigation of the role of cytochrome P4502B4 active site residues in substrate metabolism based on crystal structures of the ligand-bound enzyme.ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS,455(1),61-67. |
| MLA | Hernandez, Cynthia E.,et al."Investigation of the role of cytochrome P4502B4 active site residues in substrate metabolism based on crystal structures of the ligand-bound enzyme".ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 455.1(2006):61-67. |
入库方式： OAI收割
来源：上海药物研究所
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