Reassessment of the active site of the amine oxidase AO-I from Aspergillus niger AKU 3302 and a structure of the coding gene

Kód výsledku v IS VaVaI

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Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Reassessment of the active site of the amine oxidase AO-I from Aspergillus niger AKU 3302 and a structure of the coding gene

Popis výsledku v původním jazyce

Amine oxidase AO-I from Aspergillus niger AKU 3302 has been reported to contain topa quinone as the cofactor, however, the study on p-nitrophenylhydrazine derivatised enzyme and purified active site peptides showed the presence of a carboxylate ester linkage of topa to a glutamate [1]. Recently the catalytic functionality of such a crosslinked cofactor has been shown to be unlikely by spectroscopic and voltammetric studies on synthesised model compounds [2]. We have obtained resonance Raman spectra of the AO-I in native state and after reduction with substrate that indeed show that the catalytically active cofactor is unmodified topa quinone. The primary structure of the enzyme [3] (GenBank U31869) has been reviewed by sequencing the AO-I cDNA on a capillary DNA sequencer and shown to contain several frame shifts and errors that account for previously reported lower homology with other amine oxidases in the regions around copper binding histidine residues. The sequence was verified by

Název v anglickém jazyce

Reassessment of the active site of the amine oxidase AO-I from Aspergillus niger AKU 3302 and a structure of the coding gene

Popis výsledku anglicky

Amine oxidase AO-I from Aspergillus niger AKU 3302 has been reported to contain topa quinone as the cofactor, however, the study on p-nitrophenylhydrazine derivatised enzyme and purified active site peptides showed the presence of a carboxylate ester linkage of topa to a glutamate [1]. Recently the catalytic functionality of such a crosslinked cofactor has been shown to be unlikely by spectroscopic and voltammetric studies on synthesised model compounds [2]. We have obtained resonance Raman spectra of the AO-I in native state and after reduction with substrate that indeed show that the catalytically active cofactor is unmodified topa quinone. The primary structure of the enzyme [3] (GenBank U31869) has been reviewed by sequencing the AO-I cDNA on a capillary DNA sequencer and shown to contain several frame shifts and errors that account for previously reported lower homology with other amine oxidases in the regions around copper binding histidine residues. The sequence was verified by

Druh

D - Stať ve sborníku

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

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Návaznosti

Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2002

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název statě ve sborníku

3rd International Symposium on Vitamin B6, PQQ, Carbonyl Catalysis and quinoproteins

ISBN

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ISSN

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e-ISSN

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Počet stran výsledku

60

Strana od-do

56

Název nakladatele

University of Southampton

Místo vydání

Southampton

Místo konání akce

Southampton

Datum konání akce

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Typ akce podle státní příslušnosti

WRD - Celosvětová akce

Kód UT WoS článku

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