PURIFICATION, CHARACTERIZATION, AND COMPLETE AMINO-ACID-SEQUENCE OF A THIOREDOXIN FROM A GREEN-ALGA, CHLAMYDOMONAS-REINHARDTII

被引:41
作者
DECOTTIGNIES, P
SCHMITTER, JM
JACQUOT, JP
DUTKA, S
PICAUD, A
GADAL, P
机构
[1] LAB BIOCHIM FONCTIONNELLE MEMBRANES VEGETALES, F-91198 GIF SUR YVETTE, FRANCE
[2] ECOLE POLYTECH, BIOCHIM LAB, F-91128 PALAISEAU, FRANCE
关键词
D O I
10.1016/0003-9861(90)90525-4
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
Two thioredoxins (named Ch1 and Ch2 in reference to their elution pattern on an anion-exchange column) have been purified to homogeneity from the green alga, Chlamydomonas reinhardtii. In this paper, we describe the properties and the sequence of the most abundant form, Ch2. Its activity in various enzymatic assays has been compared with those of Escherichia coli and spinach thioredoxins. C. reinhardtii thioredoxin Ch2 can serve as a substrate for E. coli thioredoxin reductase, with a lower efficiency when compared to the homologous system. In the presence of dithiothreitol (DTT), the protein is able to catalyze the reduction of porcine insulin. Thioredoxin Ch2 is as efficient as its spinach counterpart in the DTT or light activation of corn NADP-malate dehydrogenase, but it only activates spinach fructose-1, 6-bisphosphatase at very high concentrations. The complete primary structure of the C. reinhardtii thioredoxin Ch2 was determined by automated Edman degradation of the intact protein and of peptides derived from trypsin, chymotrypsin, clostripain, and SV8 protease digestions. It consists of a polypeptide of 106 amino acids (MW 11,808) and contains the well-conserved active site sequence Trp-Cys-Gly-Pro-Cys. The sequence of the algal thioredoxin Ch2 has been compared to that of thioredoxins from other sources and has the greatest similarity (67%) with the thioredoxin from Anabaena 7119. © 1990.
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页码:112 / 121
页数:10
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