Synthesis and characterization of an immobilized phenylethanolamine N-methyltransferase liquid chromatographic stationary phase.
Norepinephrine is N-methylated to epinephrine by the catalytic effect of the terminal enzyme in catecholamine biosynthesis, phenylethanolamine N-methyltransferase (PNMT). PNMT has been covalently immobilized onto a silica-based liquid chromatographic support, glutaraldehyde-P (Glut-P). The resulting PNMT-Glut-P stationary phase (PNMT-SP) was enzymatically active, stable, and reusable. Standard Michaelis-Menten kinetic studies were performed with both free and immobilized PNMT and known substrates and inhibitors were examined. The results demonstrate that the PNMT-SP can be utilized for the rapid screening of potential PNMT substrates as well as the screening of compounds for PNMT inhibitory activity.
Markoglou
N
N
Department of Medicine, Division of Experimental Medicine, McGill University, Montreal, Quebec, Canada.
Wainer
I W
IW
D000818:Animals; D001596:Benzylamines / Q000494:pharmacology; D002853:Chromatography, Liquid / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005033:Ethylmaleimide / Q000494:pharmacology; D005976:Glutaral / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009647:Normetanephrine / Q000378:metabolism; D009941:Organomercury Compounds / Q000494:pharmacology; D010625:Phenylethanolamine N-Methyltransferase / Q000737:chemistry* / Q000378:metabolism*; D012436:S-Adenosylmethionine / Q000378:metabolism; D013696:Temperature
C400327:4-CMPS; D001596:Benzylamines; D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D009941:Organomercury Compounds; D009647:Normetanephrine; D012436:S-Adenosylmethionine; C030796:benzylamine; D010625:Phenylethanolamine N-Methyltransferase; D005033:Ethylmaleimide; D005976:Glutaral