Biochemical characterization of thioredoxin 1 from Dictyostelium discoideum.
Multiple genes for thioredoxins (TRX) have been demonstrated in Dictyostelium discoideum. We expressed the cDNA for one of these genes (DdTrx1) in E. coli and purified the protein to homogeneity. The interaction with classic substrates as well as TRX reductases was analysed. It reacted with every tested substrate: insulin, NADP-dependent malate dehydrogenase and fructose-1,6-bisphosphatase. With a S0.5 of 20 microM, the reactivity with the fructose-1,6-bisphosphatase is the highest ever found for a heterologous TRX. DdTRX1 itself is accepted as a substrate by the chloroplast ferredoxin-dependent TRX reductase, as well as by the E. coli NADPH-dependent TRX reductase. Thus, the Dictyostelium TRX is functionally promiscuous. Its reactivity with insulin, chloroplast NADP-dependent malate dehydrogenase and ferredoxin-dependent TRX reductase resemble those of other TRX. It is, however, clearly different in its good interaction with chloroplast fructose-1,6-bisphosphatase and in its poor interaction with E. coli NADP-dependent TRX reductase.
Wetterauer
B
B
Unité de Biochimie Cellulaire, Institut Pasteur, Paris, France.
Véron
M
M
Miginiac-Maslow
M
M
Decottignies
P
P
Jacquot
J P
JP
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004023:Dictyostelium / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D006597:Fructose-Bisphosphatase / Q000378:metabolism; D005800:Genes, Fungal; D007328:Insulin / Q000378:metabolism; D007700:Kinetics; D008291:Malate Dehydrogenase / Q000378:metabolism; D005810:Multigene Family; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013880:Thioredoxin-Disulfide Reductase / Q000378:metabolism; D013879:Thioredoxins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D004271:DNA, Fungal; D007328:Insulin; D011994:Recombinant Proteins; D013879:Thioredoxins; D008291:Malate Dehydrogenase; D013880:Thioredoxin-Disulfide Reductase; D006597:Fructose-Bisphosphatase