期刊
BIOCHEMICAL PHARMACOLOGY
卷 84, 期 6, 页码 775-783出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.bcp.2012.07.002
关键词
CB 1954; Nitroreductase; SOS response; Enzyme prodrug therapy; Directed evolution
资金
- Genesis Oncology Trust
- Cancer Society of New Zealand
- Victoria University Research Fund
- Health Research Council of New Zealand [06/229, 08/103]
- Maurice Wilkins Centre for Molecular Biodiscovery PhD Scholarship
Phase I/II cancer gene therapy trials of the Escherichia coli nitroreductase NfsB in partnership with the prodrug CB1954 [5-(aziridin-1-yl)-2,4-dinitrobenzamide] have indicated that CB1954 toxicity is dose-limiting at concentrations far below the enzyme K-M. Here we report that the flavin reductase FRase I from Vibrio fischeri is also a CB1954 nitroreductase, which has a substantially lower apparent K-M than E. coli NfsB. To enhance the activity of FRase I with CB1954 we used targeted mutagenesis and an E. coli SOS reporter strain to engineer single- and multi-residue variants that possess a substantially reduced apparent K-M and an increased k(cat)/K-M relative to the wild type enzyme. In a bacteria-delivered model for enzyme prodrug therapy, the engineered FRase I variants were able to kill human colon carcinoma (HCT-116) cells at significantly lower CB1954 concentrations than wild type FRase I or E. coli NfsB. (C) 2012 Elsevier Inc. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据