Human phenylethanolamine N-methyltransferase pharmacogenomics: gene re-sequencing and functional genomics.
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.
Ji
Yuan
Y
Department of Molecular Pharmacology & Experimental Therapeutics, Mayo Clinic College of Medicine, Rochester, Minnesota, USA.
Salavaggione
Oreste E
OE
Wang
Liewei
L
Adjei
Araba A
AA
Eckloff
Bruce
B
Wieben
Eric D
ED
Weinshilboum
Richard M
RM
D000818:Animals; D044383:Black People; D015153:Blotting, Western; D004247:DNA / Q000235:genetics; D005091:Exons / Q000235:genetics; D017930:Genes, Reporter; D023281:Genomics; D005838:Genotype; D006239:Haplotypes; D006801:Humans; D066298:In Vitro Techniques; D007438:Introns / Q000235:genetics; D007700:Kinetics; D015810:Linkage Disequilibrium; D008156:Luciferases / Q000235:genetics; D010641:Phenotype; D010625:Phenylethanolamine N-Methyltransferase / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D020539:Sequence Analysis, Protein; D044465:White People; D001741:Black or African American
D004247:DNA; D008156:Luciferases; D010625:Phenylethanolamine N-Methyltransferase