Abstract
Six novel nonsynonymous nucleotide alterations were found in the cytochrome P450 1A2 gene in a Japanese population, which resulted in the following amino acid substitutions: T83M, E168Q, F186L, S212C, G299A, and T438I. These individuals were heterozygous for the amino acid substitutions. The potential functional alterations caused by the amino acid substitutions were characterized by a cDNA-mediated expression system using Chinese hamster V79 cells. Among the six CYP1A2 variants, F186L showed the most profound and statistically significant reduction in O-deethylation of phenacetin and 7-ethoxyresorufin. Kinetic analyses performed for the ethoxyresorufin O-deethylation revealed that the Vmax of the F186L variant was approximately 5% of that of the CYP1A2 wild type, despite a 5-fold lower Km value of the variant, the consequence of which was reduced enzymatic activity toward the substrate. Thus, for the first time, phenylalanine at residue 186 is suggested to be a critical amino acid for catalytic activity.
Footnotes
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This study was supported in part by the Program for Promotion of Fundamental Studies in Health Sciences (MPJ-2, MPJ-3, and MPJ-6) of the Organization for Pharmaceutical Safety and Research of Japan.
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DOI: 10.1124/jpet.103.055798.
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ABBREVIATIONS: P450, cytochrome P450; PCR, polymerase chain reaction; HPLC, high-performance liquid chromatography; SNP, single nucleotide polymorphism; HPLC, high-performance liquid chromatography; SRS, substrate recognition sequence.
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↵1 These authors contributed equally to this work.
- Received June 20, 2003.
- Accepted October 9, 2003.
- The American Society for Pharmacology and Experimental Therapeutics
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