title: A sensitive radioenzymatic assay for epinephrine forming enzymes.
abstract: Epinephrine (E) is formed in the adrenal medulla by phenylethanolamine-N-methyltransferase (PNMT), and in other tissues. Enzymes other than PNMT may be able to synthesize E, but this has been difficult to investigate because most assays do not have E as their final product. This assay produces 3H-E from norepinephrine (NE) and 3H-S-adenosylmethionine. The 3H-E is isolated on alumina, 3H-S-adenosylmethionine is precipitated and the 3H-E is suspended in diethylhexyl phosphoric acid in toluene for scintillation counting. The assay is sensitive and linear over a wide range. E was formed by most tissues tested. Brain and adrenal contained an enzyme specific for NE, but cardiac ventricle contained an enzyme that methylated both NE and dopamine. Denervated tissues in adrenal medullectomized rats contained very little NE, but still had E and E forming enzyme present. This assay detects a non-neuronal E forming enzyme with activity in vitro and in vivo.
authors.affiliation: Dept. of Medicine, UCSD Medical Center 92103.
authors.forename: M G
authors.initials: MG
authors.lastname: Ziegler
authors.forename: B
authors.initials: B
authors.lastname: Kennedy
authors.forename: H
authors.initials: H
authors.lastname: Elayan
mesh_terms: D000313:Adrenal Medulla / Q000201:enzymology; D000818:Animals; D004837:Epinephrine / Q000096:biosynthesis* / Q000302:isolation & purification; D005260:Female; D007700:Kinetics; D009928:Organ Specificity; D010625:Phenylethanolamine N-Methyltransferase / Q000032:analysis*; D011865:Radioisotope Dilution Technique; D051381:Rats; D012436:S-Adenosylmethionine / Q000302:isolation & purification; D012588:Scintillation Counting / Q000379:methods; D014316:Tritium
chemical_list: D014316:Tritium; D012436:S-Adenosylmethionine; D010625:Phenylethanolamine N-Methyltransferase; D004837:Epinephrine