Re-engineering cytochrome P450 2B11dH for enhanced metabolism of several substrates including the anti-cancer prodrugs cyclophosphamide and ifosfamide

doi:10.1016/j.abb.2006.12.021

	Re-engineering cytochrome P450 2B11dH for enhanced metabolism of several substrates including the anti-cancer prodrugs cyclophosphamide and ifosfamide
	Sun, Ling ; Chen, Chong S.; Waxman, David J.; Liu, Hong; Halpert, James R.; Kumar, Santosh
刊名	ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
	2007-02-15
卷号	458 期号:2 页码:167-174
关键词	cytochrome p450 P4502B11 P450 engineering structure-function relationships site-directed mutagenesis enzyme catalysis
ISSN号	0003-9861
DOI	10.1016/j.abb.2006.12.021
文献子类	Article
英文摘要	Based on recent directed evolution of P450 2B1, six P450 2B11 mutants at three positions were created in an N-terminal modified construct termed P450 2B11 dH and characterized for enzyme catalysis using five substrates. Mutant I209A demonstrated a 3.2-fold enhanced k(cat)/K-m for 7-ethoxy-4-trifluoromethylcourmarin O-deethylation, largely due to a dramatic decrease in K-m (0.72 mu M vs. 18 mu M). 1209A also demonstrated enhanced selectivity for testosterone 16 beta-hydroxylation over 16 alpha-hydroxylation. In contrast, V183L showed a 4-fold increased k(cat) for 7-benzyloxyresorufin debenzylation and a 4.7-fold increased k(cat)/K-m for testosterone 16a-hydroxylation. V183L also displayed a 1.7-fold higher k(cat)/K-m than P450 2B11dH with the anti-cancer prodrugs cyclophosphamide and ifosfamide, resulting from a similar to 4-fold decrease in Km. Introduction of the V183L mutation into full-length P450 21311 did not enhance the k(cat)/K-m. Overall, the re-engineered P450 2B11dH enzymes exhibited enhanced catalytic efficiency with several substrates including the anti-cancer prodrugs. (c) 2007 Elsevier Inc. All rights reserved.
资助项目	NIEHS NIH HHS[ES06676] ; NIEHS NIH HHS[R01 ES003619] ; NIEHS NIH HHS[5 P42 ES07381] ; NIEHS NIH HHS[P30 ES006676] ; NIEHS NIH HHS[ES03619] ; NIEHS NIH HHS[P42 ES007381] ; NCI NIH HHS[R01 CA049248] ; NCI NIH HHS[CA49248]
WOS关键词	SITE-DIRECTED MUTAGENESIS ; ACTIVE-SITE ; LABORATORY EVOLUTION ; 1A2 MUTANTS ; P4502B1 ; RESIDUES ; HYDROXYLATION ; IDENTIFICATION ; ENZYME ; ROLES
WOS研究方向	Biochemistry & Molecular Biology ; Biophysics
语种	英语
出版者	ELSEVIER SCIENCE INC
WOS记录号	WOS:000244385000010
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/273323]
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.Boston Univ, Dept Biol, Boston, MA 02215 USA 3.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	Sun, Ling,Chen, Chong S.,Waxman, David J.,et al. Re-engineering cytochrome P450 2B11dH for enhanced metabolism of several substrates including the anti-cancer prodrugs cyclophosphamide and ifosfamide[J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS,2007,458(2):167-174.
APA	Sun, Ling,Chen, Chong S.,Waxman, David J.,Liu, Hong,Halpert, James R.,&Kumar, Santosh.(2007).Re-engineering cytochrome P450 2B11dH for enhanced metabolism of several substrates including the anti-cancer prodrugs cyclophosphamide and ifosfamide.ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS,458(2),167-174.
MLA	Sun, Ling,et al."Re-engineering cytochrome P450 2B11dH for enhanced metabolism of several substrates including the anti-cancer prodrugs cyclophosphamide and ifosfamide".ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 458.2(2007):167-174.