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Human catechol O-methyltransferase genetic variation: gene resequencing and functional characterization of variant allozymes

Abstract

Catechol O-methyltransferase (COMT) plays an important role in the metabolism of catecholamines, catecholestrogens and catechol drugs. A common COMT G472A genetic polymorphism (Val108/158Met) that was identified previously is associated with decreased levels of enzyme activity and has been implicated as a possible risk factor for neuropsychiatric disease. We set out to ‘resequence’ the human COMT gene using DNA samples from 60 African-American and 60 Caucasian-American subjects. A total of 23 single nucleotide polymorphisms (SNPs), including a novel nonsynonymous cSNP present only in DNA from African-American subjects, and one insertion/deletion were observed. The wild type (WT) and two variant allozymes, Thr52 and Met108, were transiently expressed in COS-1 and HEK293 cells. There was no significant change in level of COMT activity for the Thr52 variant allozyme, but there was a 40% decrease in the level of activity in cells transfected with the Met108 construct. Apparent Km values of the WT and variant allozymes for the two reaction cosubstrates differed slightly, but significantly, for 3,4-dihydroxybenzoic acid but not for S-adenosyl-L-methionine. The Met108 allozyme displayed a 70–90% decrease in immunoreactive protein when compared with WT, but there was no significant change in the level of immunoreactive protein for Thr52. A significant decrease in the level of immunoreactive protein was also observed in hepatic biopsy samples from patients homozygous for the allele encoding Met108. These observations represent steps toward an understanding of molecular genetic mechanisms responsible for variation in COMT level and/or properties, variation that may contribute to the pathophysiology of neuropsychiatric disease.

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Acknowledgements

We thank Dr Daniel Schaid for performing the linkage and haplotype analyses and Ms Luanne Wussow for her assistance with the preparation of this manuscript. This work was supported in part by NIH Grants RO1 GM28157 (RMW), RO1 GM35720 (RMW), PO1 CA82267 (RWM), UO1 GM61388 (RMW and EDW) and R25T CA92049 (AJS).

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Correspondence to R M Weinshilboum.

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The DNA resequencing data described in this manuscript have been deposited in the NIH-sponsored Pharmacogenetics Research Network database PharmGKB with Accession Number PA117.

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Shield, A., Thomae, B., Eckloff, B. et al. Human catechol O-methyltransferase genetic variation: gene resequencing and functional characterization of variant allozymes. Mol Psychiatry 9, 151–160 (2004). https://doi.org/10.1038/sj.mp.4001386

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Keywords

  • catechol O-methyltransferase
  • COMT
  • methylation
  • genetic polymorphisms
  • SNPs
  • pharmacogenetics

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