摘要
目的:纯化细胞色素氧化酶P450(CYP4F3A)以及测定酶促反应动力学参数。方法:应用免疫共沉淀技术纯化CYP4F3A酶,通过LTB4降解实验测定其米氏常数。结果:纯化的CYP4F3A浓度为0.12 mg/ml,Km=2.643±0.106μmol/L,Vmax=24.97±1.163 nmol/(min.nmol)。结论:获得从哺乳动物细胞中纯化的CYP4F3A具有较强LTB4降解活性,并通过测定其酶动力学常数,为研究CYP4F3A活性调节机制打下基础。
Objective ：To purify CYP4F3A and investigate the parameters of its enzyme kinetics. Methods： Human cytochrorae P450（CYP4F3A） expression was detected by Western-blot and CYP4F3A was purified by imrauno-coprecipitation and Flag peptide substitution. The kinetic analysis of CYP4F3A was conducted by degradation of the substrate（ LTB4 ）. The LTB4 level was determined using a commercial ELISA kit. Results：Purified CYP4F3A had normal LTB4 ω-hydroxylase activity. Km = 2.643 ± 0. 106 μmol/L, Vmax = 24.97 ± 1. 163 nmol/（min · nmol）. Conclusion： CYP4F3A was successfully purified from human embryo kidney 293T cells and its kinetic constant was determined.
出处
《军事医学科学院院刊》
CSCD
北大核心
2009年第3期234-236,共3页
Bulletin of the Academy of Military Medical Sciences
基金
“863”计划重点资助项目(2005AA218080)
