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

J Neurochem. 2005 Dec;95(6):1766-76. doi: 10.1111/j.1471-4159.2005.03453.x. Epub 2005 Nov 8.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Black People
  • Black or African American
  • Blotting, Western
  • DNA / genetics
  • Exons / genetics
  • Genes, Reporter
  • Genomics
  • Genotype
  • Haplotypes
  • Humans
  • In Vitro Techniques
  • Introns / genetics
  • Kinetics
  • Linkage Disequilibrium
  • Luciferases / genetics
  • Phenotype
  • Phenylethanolamine N-Methyltransferase / genetics*
  • Polymorphism, Single Nucleotide
  • Rabbits
  • Reticulocytes / metabolism
  • Sequence Analysis, Protein
  • White People

Substances

  • DNA
  • Luciferases
  • Phenylethanolamine N-Methyltransferase