Further studies on the metabolism of methylamine by semicarbazide-sensitive amine oxidase activities in human plasma, umbilical artery and rat aorta.
An ion exchange radiochemical assay has been developed to study the deamination of [14C]methylamine (MA) in homogenates of rat aorta and human umbilical artery, as well as in samples of human plasma. MA metabolism was found to be inhibited almost completely by 1 mM semicarbazide, but virtually unaffected by 0.1 mM clorgyline, suggesting that MA is a substrate for the semicarbazide-sensitive amino oxidase (SSAO) activities which also metabolize benzylamine (BZ) in these sources. Mean Km values for MA metabolism by aorta, umbilical artery and plasma were 182, 832 and 516 microM, respectively, with corresponding Vmax values in aorta and umbilical artery of 100 and 590 nmol (mg prot.)-1 h-1, and in plasma of 48 nmol (mL serum)-1 h-1. Kinetic constants determined for [14C]BZ metabolism in plasma (by an organic solvent extraction assay) and in umbilical artery (by the ion exchange assay) yielded mean Km values of 225 microM (plasma), 222 microM (umbilical artery), and Vmax values of 28 nmol (mL serum)-1 h-1 (plasma) and 377 nmol (mg prot.)-1 h-1 (umbilical artery). The deamination of [14C]MA was inhibited competitively by unlabelled BZ, with Ki values in umbilical artery and plasma of 220 and 172 microM, respectively. Also, metabolite formation from mixtures of [14C]BZ (200 microM) and [14C]MA (800 microM) was extremely close to that predicted for a single enzyme capable of metabolizing two alternative substrates in a competitive fashion.(ABSTRACT TRUNCATED AT 250 WORDS)
Lyles
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GA
Department of Pharmacology and Clinical Pharmacology, Ninewells Hospital, Dundee, UK.
Holt
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A
Marshall
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CM
D000629:Aminopropionitrile / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic / Q000201:enzymology; D003010:Clorgyline / Q000494:pharmacology; D003124:Colorimetry; D003956:Dialysis; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008744:Methylamines / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000201:enzymology*; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000037:antagonists & inhibitors* / Q000097:blood; D011506:Proteins / Q000378:metabolism; D051381:Rats; D012663:Semicarbazides / Q000494:pharmacology*; D014469:Umbilical Arteries / Q000201:enzymology
D008744:Methylamines; D011506:Proteins; D012663:Semicarbazides; D000629:Aminopropionitrile; D000587:Oxidoreductases Acting on CH-NH Group Donors; D003010:Clorgyline