Site-directed mutagenesis around the CuA site of a polyphenol oxidase from Coreopsis grandiflora (cgAUS1)

被引：17
|
作者：
Kaintz, Cornelia [1 ]
Mayer, Rupert L. [2 ]
Jirsa, Franz [3 ]
Halbwirth, Heidi [4 ]
Rompel, Annette [1 ]
机构：
[1] Univ Wien, Fak Chem, Inst Biophys Chem, A-1090 Vienna, Austria
[2] Univ Wien, Dept Analyt Chem, A-1090 Vienna, Austria
[3] Univ Wien, Dept Inorgan Chem, A-1090 Vienna, Austria
[4] Vienna Univ Technol, Inst Chem Engn, A-1060 Vienna, Austria
来源：
FEBS LETTERS | 2015年 / 589卷 / 07期
基金：
奥地利科学基金会;
关键词：
Type-3 copper protein; Polyphenol oxidase (PPO); Aurone synthase (AUS); Site-directed mutagenesis; 4-Deoxyaurone; Copper binding site; ASPERGILLUS-ORYZAE; CRYSTAL-STRUCTURE; ESCHERICHIA-COLI; CATECHOL OXIDASE; AGARICUS-BISPORUS; TYROSINASE GENE; CLONING; EXPRESSION; FUNGAL; PLANT;
D O I：
10.1016/j.febslet.2015.02.009
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：
Aurone synthase from Coreopsis grandiflora (cgAUS1), catalyzing conversion of butein to sulfuretin in a type-3 copper center, is a rare example of a polyphenol oxidase involved in anabolism. Site-directed mutagenesis around the CuA site of AUS1 was performed, and recombinant enzymes were analyzed by mass spectrometry. Replacement of the coordinating CuA histidines with alanine resulted in the presence of a single copper and loss of diphenolase activity. The thioether bridge-building cysteine and a phenylalanine over the CuA site, exchanged to alanine, have no influence on copper content but appear to play an important role in substrate binding. (C) 2015 The Authors. Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
引用
收藏
页码：789 / 797
页数：9
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