Six novel nonsynonymous CYP1A2 gene polymorphisms: catalytic activities of the naturally occurring variant enzymes.
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.
Murayama
Norie
N
Project Team for Pharmacogenetics, National Institute of Health Sciences, Tokyo, Japan.
Soyama
Akiko
A
Saito
Yoshiro
Y
Nakajima
Yukiko
Y
Komamura
Kazuo
K
Ueno
Kazuyuki
K
Kamakura
Shiro
S
Kitakaze
Masafumi
M
Kimura
Hideo
H
Goto
Yu-ichi
Y
Saitoh
Osamu
O
Katoh
Masaaki
M
Ohnuma
Teiichi
T
Kawai
Mitsuru
M
Sugai
Kenji
K
Ohtsuki
Taisuke
T
Suzuki
Chieko
C
Minami
Narihiro
N
Ozawa
Shogo
S
Sawada
Jun-ichi
J
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D006224:Cricetinae; D019388:Cytochrome P-450 CYP1A2 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010078:Oxazines / Q000378:metabolism; D010615:Phenacetin / Q000378:metabolism; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014162:Transfection
D010078:Oxazines; C007649:ethoxyresorufin; D019388:Cytochrome P-450 CYP1A2; D010615:Phenacetin