caa a22 4500 378858939 CHVBK 20180305123341.0 cr unu---uuuuu 161128e20031107xx s 000 0 eng 10.1515/BC.2003.161 doi (NATIONALLICENCE)gruyter-10.1515/BC.2003.161 Gene Organization and Molecular Modeling of Copper Amine Oxidase from Aspergillus niger: Re-Evaluation of the Cofactor Structure [Elektronische Daten] [I. Frébort, M. Šebela, S. Hirota, M. Yamada, H. Tamaki, H. Kumagai, O. Adachi, P. Peč] Amine oxidase AO-I from Aspergillus niger AKU 3302 has been reported to contain topa quinone (TPQ) as a cofactor; however, analysis of the p-nitrophenylhydrazine-derivatized enzyme and purified active site peptides showed the presence of a carboxylate ester linkage of TPQ to a glutamate. The catalytic functionality of such a cross-linked cofactor has recently been shown unlikely by spectroscopic and voltammetric studies on synthesized model compounds. We have obtained resonance Raman spectra of native and substrate-reduced AO-I demonstrating that the catalytically active cofactor is unmodified TPQ. The primary structure of the enzyme (GenBank acc. no. U31869) has been reviewed and updated by repeated isolation and sequencing of AOI cDNA. This allowed rectification of several errors that account for previously reported low homology to other amine oxidases in the regions around copper binding histididyl residues. The results were confirmed by cloning the ao-1 structural gene (GenBank acc. no. AF362473). Analysis of the gene 5'-upstream region of the gene revealed potential binding sites for an analog of NIT2, the nitrogen metabolism regulatory protein found in Neurospora crassa and other fungi. The molecular structure of AO-I was modeled by a comparative method using published crystal structures of amine oxidases as templates. Copyright © 2003 by Walter de Gruyter GmbH & Co. KG Biochemistry nationallicence Molecular biology nationallicence Cellular biology nationallicence Frébort I. aut Šebela M. aut Hirota S. aut Yamada M. aut Tamaki H. aut Kumagai H. aut Adachi O. aut Peč P. aut Biological Chemistry Walter de Gruyter 384/10-11(2003-11-07), 1451-1461 1431-6730 384:10-11<1451 2003 384 bchm https://doi.org/10.1515/BC.2003.161 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/BC.2003.161 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Frébort I. aut NATIONALLICENCE 700 1- Šebela M. aut NATIONALLICENCE 700 1- Hirota S. aut NATIONALLICENCE 700 1- Yamada M. aut NATIONALLICENCE 700 1- Tamaki H. aut NATIONALLICENCE 700 1- Kumagai H. aut NATIONALLICENCE 700 1- Adachi O. aut NATIONALLICENCE 700 1- Peč P. aut NATIONALLICENCE 773 0- Biological Chemistry Walter de Gruyter 384/10-11(2003-11-07), 1451-1461 1431-6730 384:10-11<1451 2003 384 bchm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter