title: Reductive trapping of substrate to methylamine oxidase from Arthrobacter P1.
abstract: Methylamine oxidase (EC 1.4.3.6) from Arthrobacter P1 was inactivated by NaCNBH3 in the presence of [14C]benzylamine, leading to the incorporation of 1 mol of radiolabeled substrate/mol of enzyme subunit at complete inactivation. By contrast, no labeling of enzyme was observed using [3H]NaCNBH3 as reductant. These results are analogous to those previously reported for the eukaryotic enzyme, bovine serum plasma amine oxidase [(1987) J. Biol. Chem. 262, 962-965]. The observed pattern of labeling is consistent with the presence of dicarbonyl cofactor at the active site of methylamine oxidase. Further, these studies suggest that our reductive trapping technique, in which the pattern of radiolabeling of an enzyme is compared using C-14 substrate vs tritiated reductant, may serve as a general assay for covalently bound dicarbonyl structures.
authors.affiliation: Department of Chemistry, University of California, Berkeley 94720.
authors.forename: C
authors.initials: C
authors.lastname: Hartmann
authors.forename: J P
authors.initials: JP
authors.lastname: Klinman
mesh_terms: D001173:Arthrobacter / Q000201:enzymology*; D001596:Benzylamines / Q000378:metabolism*; D001894:Borohydrides / Q000494:pharmacology; D002250:Carbon Radioisotopes; D055598:Chemical Phenomena; D002621:Chemistry; D003956:Dialysis; D004789:Enzyme Activation / Q000187:drug effects; D007553:Isotope Labeling; D007700:Kinetics; D010084:Oxidation-Reduction; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D045542:PQQ Cofactor; D010659:Phenylhydrazines / Q000494:pharmacology; D015363:Quinolones / Q000032:analysis; D013053:Spectrophotometry; D014316:Tritium
chemical_list: D001596:Benzylamines; D001894:Borohydrides; D002250:Carbon Radioisotopes; D010659:Phenylhydrazines; D015363:Quinolones; C030299:phenylhydrazine; D014316:Tritium; D045542:PQQ Cofactor; C030796:benzylamine; C009282:sodium cyanoborohydride; C021567:methylamine dehydrogenase; D000587:Oxidoreductases Acting on CH-NH Group Donors