Probing functional interactions between cytochromes P450 with principal component analysis of substrate saturation profiles and targeted proteomics.
Probing functional interactions between cytochromes P450 with principal component analysis of substrate saturation profiles and targeted proteomics.
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探测细胞色素P450之间的功能相互作用与底物饱和度和靶向蛋白质组学的主要成分分析。
DOI：
10.1016/j.abb.2021.108937
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发表时间：
2021-09-15
影响因子：
3.9
通讯作者：
Davydov DR
中科院分区：
文献类型：
--
作者：
Dangi B;Davydova NY;Maldonado MA;Ahire D;Prasad B;Davydov DR
我们研究了药物代谢途径与人肝微粒体（HLM）中P450集合组成之间的对应关系。作为探针，我们使用香豆素152（C152），荧光底物代谢的多个P450物种。研究了7个混合HLM制剂中的底物饱和度曲线（SSP），我们试图将其与靶向蛋白质组学表征的P450池的组成相关联。该分析与使用CYP 3A 4和CYP 2C 19（主要C152代谢物）特异性抑制剂的试验相补充，证明了不同HLM样品之间的显著差异。为了揭示这些差异的来源，我们实施了主成分分析（PCA）的SSP系列与HLM样品与已知的组成的P450池。C152的代谢参数主要由CYP 2A 6、CYP 2B 6、CYP 2C 8、CYP 2 E1和CYP 3A 5的含量决定，只有CYP 2B 6和CYP 3A 5才能代谢C152。为了验证这一发现，我们研究了用CYP 2A 6、CYP 2 E1和CYP 3A 5富集HLM的效果。CYP 3A 5掺入HLM降低了C152代谢速率，同时增加了CYP 2B 6在其转换中的作用。相比之下，CYP 2A 6和CYP 2 E1的掺入使C152去甲基化转向对底物具有中等亲和力的一些P450酶，最可能是CYP 3A 4。我们的研究结果揭示了一个尖锐的非加和性的个人P450的属性，并建议一个关键的作用，P450-P450的相互作用，在确定药物代谢途径。这项研究表明，我们的新的PCA为基础的方法在揭示不同的P450物种之间的功能相互关系的高潜力。
We investigated the correspondence between drug metabolism routes and the composition of the P450 ensemble in human liver microsomes (HLM). As a probe, we used Coumarin 152 (C152), a fluorogenic substrate metabolized by multiple P450 species. Studying the substrate-saturation profiles (SSP) in seven pooled HLM preparations, we sought to correlate them with the P450 pool’s composition characterized by targeted proteomics. This analysis, complemented with the assays with specific inhibitors of CYP3A4 and CYP2C19, the primary C152 metabolizers, demonstrated a significant contrast between different HLM samples. To unveil the source of these differences, we implemented Principal Component Analysis (PCA) of the SSP series obtained with HLM samples with a known composition of the P450 pool. Our analysis revealed that the parameters of C152 metabolism are primarily determined by the content of CYP2A6, CYP2B6, CYP2C8, CYP2E1, and CYP3A5 of those only CYP2B6 and CYP3A5 can metabolize C152. To validate this finding, we studied the effect of enriching HLM with CYP2A6, CYP2E1, and CYP3A5. The incorporation of CYP3A5 into HLM decreases the rate of C152 metabolism while increasing the role of CYP2B6 in its turnover. In contrast, incorporation of CYP2A6 and CYP2E1 reroutes the C152 demethylation towards some P450 enzyme with a moderate affinity to the substrate, most likely CYP3A4. Our results reveal a sharp non-additivity of the individual P450 properties and suggest a pivotal role of P450-P450 interactions in determining drug metabolism routes. This study demonstrates the high potential of our new PCA-based approach in unveiling functional interrelationships between different P450 species.
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人类微粒体药物代谢中的非吸附性在一项研究中揭示了香豆素152（一种多元特异性的细胞色素P450底物）的研究。
DOI：
10.1080/00498254.2020.1775913
发表时间：
2020-12
期刊：
Xenobiotica; the fate of foreign compounds in biological systems
影响因子：
--
作者：
Dangi B;Davydova NY;Vavilov NE;Zgoda VG;Davydov DR
通讯作者：
Davydov DR
影响因子：
3.9
作者：
Prasad, Bhagwat;Bhatt, Deepak Kumar;Unadkat, Jashvant D.
通讯作者：
Unadkat, Jashvant D.
基因多态性和细胞色素P450含量对人肝微粒体代谢活动的影响有限
影响因子：
4.6
作者：
Gao, Na;Tian, Xin;Qiao, Hailing
通讯作者：
Qiao, Hailing
DOI：
10.1042/bcj20170543
发表时间：
2017-10-10
期刊：
The Biochemical journal
影响因子：
--
作者：
Davydov DR;Davydova NY;Rodgers JT;Rushmore TH;Jones JP
通讯作者：
Jones JP
影响因子：
13.5
作者：
Reed, James R.;Backes, Wayne L.
通讯作者：
Backes, Wayne L.