Human phenylethanolamine N-methyltransferase pharmacogenomics: Gene re-sequencing and functional genomics

Yuan Ji, Oreste E. Salavaggione, Liewei M Wang, Araba A. Adjei, Bruce Eckloff, Eric D Wieben, Richard M Weinshilboum

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Phenylethanolamine N-methyltransferase (PNMT, EC2.1.1.28) catalyzes the N-methylation of norepinephrine to form epinephrine. As a step toward understanding the possible contribution of inheritance to individual variation in PNMT-catalyzed epinephrine formation, we 're-sequenced' the entire human PNMT gene, including the three exons, the introns and approximately 1 kb of the 5′-flanking region (5′-FR), using DNA samples from 60 African-American (AA) and 60 Caucasian-American (CA) subjects. Within the 3.5 kb re-sequenced, 18 single nucleotide polymorphisms (SNPs) were observed, including four non-synonymous coding SNPs (cSNPs) that resulted in the following alterations in encoded amino acid sequence: Asn9Ser, Thr98Ala, Arg112Cys and Ala175Thr. When constructs for the non-synonymous cSNPs were transiently expressed in COS-1 cells, the Ala98 allozyme displayed significantly lower levels of both activity and immunoreactive protein (p < 0.002) than did the wild-type (WT) enzyme due, at least in part, to accelerated protein degradation by a proteasome-mediated process. Luciferase reporter gene constructs were also created for the six common PNMT 5′-FR haplotypes observed. Significant differences were observed among haplotypes in their ability to drive transcription. These observations raise the possibility of inherited variation in the ability to form epinephrine from norepinephrine as a result of variant PNMT polymorphisms and haplotypes.

Original languageEnglish (US)
Pages (from-to)1766-1776
Number of pages11
JournalJournal of Neurochemistry
Volume95
Issue number6
DOIs
StatePublished - Dec 2005

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Phenylethanolamine N-Methyltransferase
Pharmacogenetics
Genomics
Polymorphism
Haplotypes
Epinephrine
Single Nucleotide Polymorphism
Genes
5' Flanking Region
Norepinephrine
Nucleotides
Methylation
COS Cells
Proteasome Endopeptidase Complex
Transcription
Luciferases
Reporter Genes
African Americans
Introns
Isoenzymes

Keywords

  • Allozymes
  • Functional genomics
  • Gene resequencing
  • Haplotypes
  • Non-synonymous coding single nucleotide polymorphisms
  • Pharmacogenomics
  • Phenylethanolamine N-methyltransferase
  • Polymorphisms
  • Single nucleotide polymorphism

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Human phenylethanolamine N-methyltransferase pharmacogenomics : Gene re-sequencing and functional genomics. / Ji, Yuan; Salavaggione, Oreste E.; Wang, Liewei M; Adjei, Araba A.; Eckloff, Bruce; Wieben, Eric D; Weinshilboum, Richard M.

In: Journal of Neurochemistry, Vol. 95, No. 6, 12.2005, p. 1766-1776.

Research output: Contribution to journalArticle

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AU - Ji, Yuan

AU - Salavaggione, Oreste E.

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AU - Eckloff, Bruce

AU - Wieben, Eric D

AU - Weinshilboum, Richard M

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