Purification, cloning, and three-dimensional structure prediction of Micrococcus luteus FAD-containing tyramine oxidase

Jung Hyeob Roh, Johan Wouters, Eric Depiereux, Hideaki Yukawa, Masayuki Inui, Hiromichi Minami, Hideyuki Suzuki, Hidehiko Kumagai

Research output: Contribution to journalArticle

Abstract

The FAD-containing tyramine oxidase enzyme and gene from the Gram (+) bacterium Micrococcus luteus were isolated, and computer prediction was used to propose a preliminary 3D model of the protein. A 2.8-kb Sau3AI fragment containing the structural gene of tyramine oxidase was cloned from a M. luteus genomic DNA library. The 1332 bp gene encodes a protein of 443 amino acids, with a calculated molecular mass of 49.1 kDa. The enzyme was found to be a homodimer with a molecular weight of 49,000. It oxidizes tyramine, adrenaline, 3-hydroxytyramine, dopamine, and noradrenaline, and was reversibly inhibited by FAD-containing monoamine oxidase A and B specific inhibitors. Sequence comparison show that tyramine oxidase is smaller than other FAD-amine oxidases but that it contains well-conserved amino acid residues reported in all other FAD-amine oxidases. A hypothetical three-dimensional structure of tyramine oxidase has also been proposed based on secondary structure predictions, threading, and comparative modeling. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)293-297
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume268
Issue number2
DOIs
Publication statusPublished - 16 Feb 2000

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