Structural and functional insights into polymorphic enzymes of cytochrome P450 2C8

• DOI: 10.1007/s00726-010-0743-8
• Publication Date: 2010-09-16T09:27:00Z
• Authors: Hualin Jiang; Fangfang Zhong; Lu Sun; Weiyue Feng; Zhong-Xian Huang; Xiangshi Tan
• Keywords: Models, Molecular; 0301 basic medicine; Binding Sites; Polymorphism, Genetic; Paclitaxel; Gene Expression; Quantitative Structure-Activity Relationship; Antineoplastic Agents; Heme; Recombinant Proteins; 3. Good health; Cytochrome P-450 CYP2C8; Isoenzymes; Kinetics; 03 medical and health sciences; Catalytic Domain; Escherichia coli; Microsomes, Liver; Humans; Metabolic Detoxication, Phase I; Aryl Hydrocarbon Hydroxylases; Precision Medicine; Protein Binding

The cytochrome P450 (CYP) superfamily plays a key role in the oxidative metabolism of a wide range of drugs and exogenous chemicals. CYP2C8 is the principal enzyme responsible for the metabolism of the anti-cancer drug paclitaxel in the human liver. Nearly all previous works about polymorphic variants of CYP2C8 were focused on unpurified proteins, either cells or human liver microsomes; therefore their structure-function relationships were unclear. In this study, two polymorphic enzymes of CYP2C8 (CYP2C8.4 (I264M) and CYP2C8 P404A) were expressed in E. coli and purified. Metabolic activities of paclitaxel by the two purified polymorphic enzymes were observed. The activity of CYP2C8.4 was 25% and CYP2C8 P404A was 30% of that of WT CYP2C8, respectively. Their structure-function relationships were systematically investigated for the first time. Paclitaxel binding ability of CYP2C8.4 increased about two times while CYP2C8 P404A decreased about two times than that of WT CYP2C8. The two polymorphic mutant sites of I264 and P404, located far from active site and substrate binding sites, significantly affect heme and/or substrate binding. This study indicated that two important nonsubstrate recognition site (SRS) residues of CYP2C8 are closely related to heme binding and/or substrate binding. This discovery could be valuable for explaining clinically individual differences in the metabolism of drugs and provides instructed information for individualized medication.