Reaction pathway of bovine aortic lysyl oxidase.
The catalysis of amine oxidation by lysyl oxidase has been probed to assess for the likely order of substrate binding and product release and to discriminate between mechanistic alternatives previously proposed for other copper-dependent amine oxidases using molecular oxygen as a substrate. Lineweaver-Burk plots revealed a pattern of parallel lines when the oxidation of n-butylamine was followed at different fixed concentrations of oxygen consistent with a "ping-pong" kinetic mechanism in which the aldehyde is produced and released before the binding of oxygen, the second substrate. Initial burst experiments revealed the ability of lysyl oxidase to form and release n-butyraldehyde in amounts stoichiometric with functional active site content in the absence of oxygen, consistent with the ping-pong kinetics obtained. Reciprocal plots of n-butylamine oxidation in the presence of fixed concentrations of the reaction products were consistent with a Uni Uni Uni Bi ping-pong kinetic mechanism with the aldehyde being the first, H2O2 the second, and ammonia the last departing product. Moreover, spectral studies of the oxidation of p-hydroxybenzylamine by lysyl oxidase indicated that the enzyme does not process the amine substrate to a noncovalently bound p-hydroxybenzaldimine intermediate subsequently to be hydrolyzed to p-hydroxybenzaldehyde. The kinetic mechanism of lysyl oxidase thus appears to be similar to those described for diamine oxidase and pig plasma monoamine oxidase.
Williamson
P R
PR
Kagan
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HM
D000447:Aldehydes / Q000378:metabolism; D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D000693:Anaerobiosis; D000818:Animals; D001011:Aorta / Q000201:enzymology*; D001596:Benzylamines / Q000378:metabolism; D002082:Butylamines / Q000378:metabolism; D002417:Cattle; D007097:Imines / Q000378:metabolism; D007700:Kinetics; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D008249:Protein-Lysine 6-Oxidase / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000447:Aldehydes; D001596:Benzylamines; D002082:Butylamines; D007097:Imines; C048829:4-hydroxybenzylamine; D000594:Amino Acid Oxidoreductases; D008249:Protein-Lysine 6-Oxidase; C018475:butyraldehyde; C032462:n-butylamine; D010100:Oxygen