Subject Area物理化学
Evaluation of the two Mutagenesis of Cytochrome P450-3A4 in Normal Chinese Han Population in Shanghai Area
Cheng J(程婕); Liu Y(刘勇); Wan DF(万大方); Gu JR(顾建人); Yang L(杨凌)
Conference NameThe 1st asia Pacific ISSX Meeting
Conference Date2006-5-24
Conference Place韩国
AbstractObjective: To study the effect of the mutagenesis of C43T and T90C in gene AY606313 (discovered in normal Chinese Han population in Shanghai area) on the enzyme activity of cytochrome P450-3A4 using novel yeast expression system BWG-CPR/3A4 (published by Protein Expression and Purification, 2006). Methods: Using the gene of AY606313 (AY) as the templet, the mutants CYP3A4 AY-43 (single mutagenesis at 43bp), CYP3A4 AY-90 (single mutagenesis at 90bp), and the wild type CYP3A4 (W, the same sequence with NM017460) were obtained by PCR amplification. The genes of CYP3A4 AY, AY-43, AY-90 and W were inserted into the expression vector respectively and co-expressed with human cytochrome P450 reductase in BWG-CPR/3A4 system. Enzyme kinetics for testosterone and nifedipine metabolism have been determined for mutants and wild-type CYP3A4. Results: The activities of mutants and wild type CYP3A4 for testosterone and nifedipine metabolism were similar, the values of S50 and Vmax for the formation of 6β-OH hydroxylation testosterone were from 19.5 uM to 26.3 uM and from 7.5 pmol protein/min/pmol to 8.1 pmol protein/min/pmol, and S50 and Vmax for nifedipine oxidase activity were from 4.0 uM to 5.0 uM and from 6.9 pmol protein/min/pmol to 8.3 pmol protein/min/pmol respectively. The Ki for ketoconazole, a CYP3A4 selective inhibitor, were unchanged by CYP3A4 AY, AY-43 and AY-90, compared with that of the wild type CYP3A4. Conclusions: The results proposed that the two mutagenesis of N-terminal CYP3A4 may have less effect on the activity and expression level of CYP3A4 in this system; thereby the two N-terminal mutants may locate out of the catalytic center of CYP3A4, and have no key role on the maintaining monooxygenase activity. (Supported by the 973 program 2003CB716006 of the Ministry of Science & Technology of China)
WOS IDWOS:000242406900156
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Document Type会议论文
Corresponding AuthorYang L(杨凌)
Recommended Citation
GB/T 7714
Cheng J,Liu Y,Wan DF,et al. Evaluation of the two Mutagenesis of Cytochrome P450-3A4 in Normal Chinese Han Population in Shanghai Area[C],2006:216/1.
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