CYP2D6.10 present in human liver microsomes shows low catalytic activity and thermal stability

Katsunori Nakamura, Noritaka Ariyoshi, Tsuyoshi Yokoi, Satoru Ohgiya, Michihiro Chida, Kazuo Nagashima, Kazuaki Inoue, Takao Kodama, Noriaki Shimada, Tetsuya Kamataki

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Comparing bufuralol 1′-hydroxylase activity among liver microsomes prepared from individuals whose CYP2D6 genotypes had been determined we found that the activity tended to decrease depending on the number of the CYP2D6*10 allele. Pre-incubation of liver microsomes from individuals homozygous for the CYP2D6*10 allele resulted in a decrease in the enzyme activity more rapidly than those from individuals homozygous for the CYP2D6*1 suggesting that not only the catalytic activity but also the thermal stability of the enzyme appeared to be affected by the genetic polymorphism. To confirm this hypothesis the kinetic parameters of CYP2D6.1 and CYP2D6.10 were compared for bufuralol 1′-hydroxylation and dextromethorphan O-demethylation using microsomes prepared from yeast transformed with plasmids carrying CYP2D6 cDNAs (*1A and *10B). Kinetic studies of these CYP2D6 forms indicated clear differences in the metabolic activities between the wild (CYP2D6.1) and the mutant enzymes (CYP2D6.10). Bufuralol 1′-hydroxylase activity in microsomes of yeast expressing CYP2D6.10 was rapidly decreased by heat treatment supporting the idea that the thermal stability of the enzyme was reduced by amino acid replacement from Pro (CYP2D6.1) to Ser (CYP2D6.10). These data strongly suggest that the thermal instability together with the reduced intrinsic clearance of CYP2D6.10 is one of the causes responsible for the known fact that Orientals show lower metabolic activities than Caucasians for drugs metabolized mainly by CYP2D6 because of a high frequency of CYP2D6*10 in Orientals.

Original languageEnglish
Pages (from-to)969-973
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume293
Issue number3
DOIs
Publication statusPublished - 2002
Externally publishedYes

Fingerprint

Cytochrome P-450 CYP2D6
Liver Microsomes
Liver
Catalyst activity
Thermodynamic stability
Hot Temperature
Enzyme Stability
Microsomes
Yeast
Enzymes
Yeasts
Alleles
Dextromethorphan
Hydroxylation
Enzyme activity
Genetic Polymorphisms
Polymorphism
Kinetic parameters
Plasmids
Complementary DNA

Keywords

  • Cytochrome P450
  • Proline-rich region
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

CYP2D6.10 present in human liver microsomes shows low catalytic activity and thermal stability. / Nakamura, Katsunori; Ariyoshi, Noritaka; Yokoi, Tsuyoshi; Ohgiya, Satoru; Chida, Michihiro; Nagashima, Kazuo; Inoue, Kazuaki; Kodama, Takao; Shimada, Noriaki; Kamataki, Tetsuya.

In: Biochemical and Biophysical Research Communications, Vol. 293, No. 3, 2002, p. 969-973.

Research output: Contribution to journalArticle

Nakamura, K, Ariyoshi, N, Yokoi, T, Ohgiya, S, Chida, M, Nagashima, K, Inoue, K, Kodama, T, Shimada, N & Kamataki, T 2002, 'CYP2D6.10 present in human liver microsomes shows low catalytic activity and thermal stability', Biochemical and Biophysical Research Communications, vol. 293, no. 3, pp. 969-973. https://doi.org/10.1016/S0006-291X(02)00328-5
Nakamura, Katsunori ; Ariyoshi, Noritaka ; Yokoi, Tsuyoshi ; Ohgiya, Satoru ; Chida, Michihiro ; Nagashima, Kazuo ; Inoue, Kazuaki ; Kodama, Takao ; Shimada, Noriaki ; Kamataki, Tetsuya. / CYP2D6.10 present in human liver microsomes shows low catalytic activity and thermal stability. In: Biochemical and Biophysical Research Communications. 2002 ; Vol. 293, No. 3. pp. 969-973.
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T1 - CYP2D6.10 present in human liver microsomes shows low catalytic activity and thermal stability

AU - Nakamura, Katsunori

AU - Ariyoshi, Noritaka

AU - Yokoi, Tsuyoshi

AU - Ohgiya, Satoru

AU - Chida, Michihiro

AU - Nagashima, Kazuo

AU - Inoue, Kazuaki

AU - Kodama, Takao

AU - Shimada, Noriaki

AU - Kamataki, Tetsuya

PY - 2002

Y1 - 2002

N2 - Comparing bufuralol 1′-hydroxylase activity among liver microsomes prepared from individuals whose CYP2D6 genotypes had been determined we found that the activity tended to decrease depending on the number of the CYP2D6*10 allele. Pre-incubation of liver microsomes from individuals homozygous for the CYP2D6*10 allele resulted in a decrease in the enzyme activity more rapidly than those from individuals homozygous for the CYP2D6*1 suggesting that not only the catalytic activity but also the thermal stability of the enzyme appeared to be affected by the genetic polymorphism. To confirm this hypothesis the kinetic parameters of CYP2D6.1 and CYP2D6.10 were compared for bufuralol 1′-hydroxylation and dextromethorphan O-demethylation using microsomes prepared from yeast transformed with plasmids carrying CYP2D6 cDNAs (*1A and *10B). Kinetic studies of these CYP2D6 forms indicated clear differences in the metabolic activities between the wild (CYP2D6.1) and the mutant enzymes (CYP2D6.10). Bufuralol 1′-hydroxylase activity in microsomes of yeast expressing CYP2D6.10 was rapidly decreased by heat treatment supporting the idea that the thermal stability of the enzyme was reduced by amino acid replacement from Pro (CYP2D6.1) to Ser (CYP2D6.10). These data strongly suggest that the thermal instability together with the reduced intrinsic clearance of CYP2D6.10 is one of the causes responsible for the known fact that Orientals show lower metabolic activities than Caucasians for drugs metabolized mainly by CYP2D6 because of a high frequency of CYP2D6*10 in Orientals.

AB - Comparing bufuralol 1′-hydroxylase activity among liver microsomes prepared from individuals whose CYP2D6 genotypes had been determined we found that the activity tended to decrease depending on the number of the CYP2D6*10 allele. Pre-incubation of liver microsomes from individuals homozygous for the CYP2D6*10 allele resulted in a decrease in the enzyme activity more rapidly than those from individuals homozygous for the CYP2D6*1 suggesting that not only the catalytic activity but also the thermal stability of the enzyme appeared to be affected by the genetic polymorphism. To confirm this hypothesis the kinetic parameters of CYP2D6.1 and CYP2D6.10 were compared for bufuralol 1′-hydroxylation and dextromethorphan O-demethylation using microsomes prepared from yeast transformed with plasmids carrying CYP2D6 cDNAs (*1A and *10B). Kinetic studies of these CYP2D6 forms indicated clear differences in the metabolic activities between the wild (CYP2D6.1) and the mutant enzymes (CYP2D6.10). Bufuralol 1′-hydroxylase activity in microsomes of yeast expressing CYP2D6.10 was rapidly decreased by heat treatment supporting the idea that the thermal stability of the enzyme was reduced by amino acid replacement from Pro (CYP2D6.1) to Ser (CYP2D6.10). These data strongly suggest that the thermal instability together with the reduced intrinsic clearance of CYP2D6.10 is one of the causes responsible for the known fact that Orientals show lower metabolic activities than Caucasians for drugs metabolized mainly by CYP2D6 because of a high frequency of CYP2D6*10 in Orientals.

KW - Cytochrome P450

KW - Proline-rich region

KW - Saccharomyces cerevisiae

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