Modulation of CYP2C9 activity and hydrogen peroxide production by cytochrome b(5).
Modulation of CYP2C9 activity and hydrogen peroxide production by cytochrome b(5).
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DOI：
10.1038/s41598-020-72284-0
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发表时间：
2020-09-23
影响因子：
4.6
通讯作者：
Gutierrez-Merino C
中科院分区：
文献类型：
--
作者：
Gómez-Tabales J;García-Martín E;Agúndez JAG;Gutierrez-Merino C
细胞色素P450（CYP450）在药物解毒中起主要作用，细胞色素b5（cyt b5）刺激单氧化和解毒反应的催化循环。这种催化循环的附带反应可导致有毒活性氧物质（ROS）的显著产生。人类肝脏中最丰富的CYP亚型之一是CYP 2C 9，它催化包括非甾体抗炎药在内的多种药物的代谢降解。我们研究了微粒体膜结合和可溶性细胞色素b5对水杨酸羟基化为龙胆酸的调节，以及人肝微粒体（HLM）和CYP2C9杆状体中CYP2C9活性对ROS释放的调节。在常规阿司匹林剂量后达到的浓度下，CYP 2C 9占HLM中水杨酸羟基化的近75%。抗cyt b5抗体SC9513在很大程度上抑制HLM和CYP2C9杆状体中CYP2C9对水杨酸羟基化的速率，使KM增加约3倍。此外，可溶性人重组细胞色素b5刺激Vmax接近两倍，而它降低了近三倍的Km值在CYP2C9杆状体。对于NADPH依赖性ROS的产生，可溶性重组cyt b5在HLM和CYP2C9杆状体中均为强效抑制剂，抑制常数分别为1.04 ± 0.25和0.53 ± 0.06 µM cyt b5。这项研究表明，细胞色素b5的变异性可能是CYP2C9底物代谢个体间变异性的主要因素。
Cytochromes P450 (CYP) play a major role in drug detoxification, and cytochrome b5 (cyt b5) stimulates the catalytic cycle of mono-oxygenation and detoxification reactions. Collateral reactions of this catalytic cycle can lead to a significant production of toxic reactive oxygen species (ROS). One of the most abundant CYP isoforms in the human liver is CYP2C9, which catalyzes the metabolic degradation of several drugs including nonsteroidal anti-inflammatory drugs. We studied modulation by microsomal membrane-bound and soluble cyt b5 of the hydroxylation of salicylic acid to gentisic acid and ROS release by CYP2C9 activity in human liver microsomes (HLMs) and by CYP2C9 baculosomes. CYP2C9 accounts for nearly 75% of salicylic acid hydroxylation in HLMs at concentrations reached after usual aspirin doses. The anti-cyt b5 antibody SC9513 largely inhibits the rate of salicylic acid hydroxylation by CYP2C9 in HLMs and CYP2C9 baculosomes, increasing the KM approximately threefold. Besides, soluble human recombinant cyt b5 stimulates the Vmax nearly twofold while it decreases nearly threefold the Km value in CYP2C9 baculosomes. Regarding NADPH-dependent ROS production, soluble recombinant cyt b5 is a potent inhibitor both in HLMs and in CYP2C9 baculosomes, with inhibition constants of 1.04 ± 0.25 and 0.53 ± 0.06 µM cyt b5, respectively. This study indicates that variability in cyt b5 might be a major factor underlying interindividual variability in the metabolism of CYP2C9 substrates.
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影响因子：
3.6
作者：
Henderson, Colin J.;McLaughlin, Lesley A.;Wolf, C. Roland
通讯作者：
Wolf, C. Roland
影响因子：
3.6
作者：
Henderson, Colin J.;McLaughlin, Lesley A.;Wolf, C. Roland
通讯作者：
Wolf, C. Roland
影响因子：
5.8
作者：
CANOVADAVIS, E;CHIANG, JYL;WASKELL, L
通讯作者：
WASKELL, L
影响因子：
4.8
作者：
Frank, Daniel J.;Denisov, Ilia G.;Sligar, Stephen G.
通讯作者：
Sligar, Stephen G.
影响因子：
2.3
作者：
Bhattacharyya, Sudip;Sinha, Krishnendu;Sil, Parames C.
通讯作者：
Sil, Parames C.