Cloning of human 3-hydroxyanthranilic acid dioxygenase in Escherichia coli:: characterisation of the purified enzyme and its in vitro inhibition by Zn2+

被引：15

作者：

Calderone, V

Trabucco, M

Menin, V

Negro, A

Zanotti, G

机构：

[1] Univ Padua, Dept Organ Chem, I-35131 Padua, Italy

[2] Univ Padua, Biopolymer Res Ctr, I-35131 Padua, Italy

[3] Univ Padua, CRIBI, I-35131 Padua, Italy

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY | 2002年 / 1596卷 / 02期

关键词：

3-hydroxyanthranilic acid oxygenase; zinc; quinolinic acid; neurodegenerative disease;

D O I：

10.1016/S0167-4838(02)00216-9

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

3-Hydroxyanthranilic acid oxygenase (3-HAO) catalyses the conversion of 3-hydroxyanthranilic acid to quinolinic acid. Because of the involvement of quinolinic acid in the initiation of neurodegenerative phenomena, we have cloned human 3-HAO in Escherichia coli, overexpressed and purified it with the aim of studying its enzymatic activity and for future structural studies. The recombinant human protein, obtained in E. coli, retains its enzymatic activity which can occur only in the presence of Fe(II); several other metals have been tested but in no case the formation of the product has been observed. On the contrary, two of the ions tested inhibit the catalytic reaction and one of them, Zn2+, could be of physiological relevance. A circular dichroism analysis has also been performed, showing that the secondary structure is mainly of the beta type, with a minority of alpha. (C) 2002 Elsevier Science B.V. All rights reserved.

引用

页码：283 / 292

页数：10

共 29 条

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