Gene expression and characterization of a stress-induced tyrosine decarboxylase from Arabidopsis thaliana

被引:69
|
作者
Lehmann, Thomas [1 ]
Pollmann, Stephan [1 ]
机构
[1] Ruhr Univ Bochum, Dept Plant Physiol, D-44801 Bochum, Germany
关键词
Aromatic L-amino acid decarboxylase; Gene expression; Heterologous expression; Intracellular localization; Substrate specificity; Tyrosine decarboxylase; CELL SUSPENSION-CULTURES; TISSUE; ACID; BIOSYNTHESIS; LOCALIZATION; PCR;
D O I
10.1016/j.febslet.2009.05.017
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
Full-length tyrosine decarboxylase cDNA (TyrDC) from Arabidopsis thaliana was identified by rapid amplification of cDNA ends-PCR and isolated by RT-PCR. The TyrDC mRNA was substantially induced by drought stress and wounding, and was considerably decreased by salt stress. By using TyrDC protein fusions with green fluorescent protein, an intracellular localization to the cytoplasm was shown. Recombinant (His) 6-TyrDC was expressed in Escherichia coli and enzymatically characterized: it exclusively catalyzed the conversion of L-tyrosine to tyramine, exhibited an optimum temperature of 50 degrees C, and an optimum pH at approximately 8.5-9. Recombinant TyrDC protein formed tetramers, as shown by blue native gel electrophoresis. Structured summary: MINT-7040408: TyrDC (uniprotkb: Q8RY79) and TyrDC (uniprotkb: Q8RY79) bind (MI: 0407) by blue native page (MI: 0276) (C) 2009 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:1895 / 1900
页数:6
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