Identification of the quinone cofactor in mammalian semicarbazide-sensitive amine oxidase

Biochemistry
A HoltM M Palcic

Abstract

Mammalian semicarbazide-sensitive amine oxidase (SSAO) enzymes have been classified as EC 1.4.3.6 [amine:oxygen oxidoreductase (deaminating)(copper-containing)]. However, both the identity of the quinone cofactor and the presence of copper remain unconfirmed, and SSAO has proved impossible to purify to homogeneity in sufficient yield to permit cofactor identification. To circumvent this problem, we have partially purified SSAO enzymes from bovine and porcine aortae and have established, with a redox-cycling assay, that no other quinoproteins were present in enzyme preparations. Enzymes were then derivatized with (p-nitrophenyl)hydrazine (p-NPH), which forms a covalent yellow complex with the quinone cofactor. Visible absorbance spectra of derivatized bovine and porcine enzymes (respective lambdamax values 456 and 476 nm at neutral pH, shifting to 580 and 584 nm in 2 M KOH) were consistent with the presence of (2,4,5-trihydroxyphenyl)alanine quinone (TPQ) as cofactor. Resonance Raman spectra were essentially identical to that for pea seedling amine oxidase, a known TPQ-containing enzyme. Extensive digestion of SSAO enzymes, and of porcine kidney diamine oxidase, with pronase E yielded species with identical chromophoric properti...Continue Reading

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Citations

Sep 24, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hui LiYuan Zhang
Dec 31, 2003·Neurotoxicology·Jeff O'SullivanKeith F Tipton
Oct 26, 2000·Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP·F BoomsmaA H van den Meiracker
Apr 11, 2003·Fundamental & Clinical Pharmacology·P ClaudR Laine
Jun 17, 2011·Expert Opinion on Therapeutic Patents·Petra DunkelPéter Mátyus
Nov 1, 2013·American Journal of Physiology. Renal Physiology·May Y W WongMuh Geot Wong
Aug 6, 2005·The Journal of Pharmacology and Experimental Therapeutics·Luisa M Salter-CidMatthew D Linnik
May 7, 2004·The Journal of Pharmacology and Experimental Therapeutics·Diana Gubisne-HaberlePeter H Yu
Dec 1, 1999·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Gunnar Houen
Feb 23, 2019·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Michael J ReidTerry Podoll

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