Identification and characterization of a third thioredoxin h in poplar

被引:24
作者
Gelhaye, E [1 ]
Rouhier, N
Vlamis-Gardikas, A
Girardet, JM
Sautière, PE
Sayzet, M
Martin, F
Jacquot, JP
机构
[1] Univ Nancy 1, UMR 1136, Fac Sci, INRA,Interact Arbres Micro Organismes, F-54506 Vandoeuvre Les Nancy, France
[2] Karolinska Inst, Med Nobel Inst Biochem, Dept Med Biochem & Biophys, S-17177 Stockholm, Sweden
[3] Univ Nancy 1, Fac Sci, Lab Biosci UC885, INRA, F-54506 Vandoeuvre Les Nancy, France
[4] Univ Sci & Tech Lille Flandres Artois, Lab Endocrinol Annelides, UPRESA 8017, F-59655 Villeneuve Dascq, France
关键词
populus; redox regulation; recombinant; thioredoxin h;
D O I
10.1016/S0981-9428(03)00063-9
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Three full-length sequences encoding thioredoxin It have been isolated in a leaf/root of Populus trichocarpa cv. Trichobel expressed sequence tag (EST) library. One of these, popCXXS1 exhibits the nontypical active site CXXS homologous to atCXXS1. The second one, named popTrxh4, is related to at Trxh9 which forms with several other plant thioredoxin It a distinct subgroup of thioredoxins h. The third one, named popTrxh3. displays 66% identity and also a high degree of homology (81%) vs. the previously described popTrxh1. Nevertheless, the active sites of both proteins differ, since the active site of popTrxh1 (WCPPC) is a variant of the canonical WCGPC found in popTrxh3. The cDNA sequence of popTrxh3 has been introduced in an expression plasmid (pET3d) in order to express the corresponding recombinant polypeptide. The protein has been expressed to a high level, purified from Escherichia coli cells with a high yield and its catalytic properties compared to popTrxh1. Furthermore, two mutants, popTrxh1P40G and popTrxh3G41P, have been engineered in order to explore the importance of the active site residues in the thioredoxin h catalytic properties. The results are discussed in relation with known biochemical properties of thioredoxins h. (C) 2003 Editions scientifiques et medicales Elsevier SAS. All rights reserved.
引用
收藏
页码:629 / 635
页数:7
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