title: Induction by different thioredoxins of ATPase activity in coupling factor 1 from spinach chloroplasts.
abstract: ATPase activity of the coupling factor 1, CF1, isolated from spinach chloroplasts, was enhanced by reduction with dithiothreitol. Reduced thioredoxins from spinach chloroplasts, Escherichia coli and human lymphocytes replaced dithiothreitol as reductant and activator of the ATPase. CF1 must be in an oxidized activated state to be further activated by reduced thioredoxin. This state was obtained either by heating CF1 or removing the inhibitory intrinsic epsilon subunit from CF1. Efficiency and primary structure of the different thioredoxins were compared. The progressive addition of KCl during ATPase activation by reduced thioredoxin increases then decreases this process. We proposed that three basic amino acids corresponding to arginine 73 and lysines 82 and 96 in Escherichia coli thioredoxin play an important role in the anchorage of the thioredoxin to the negatively charged surface of the CF1 and are involved in the dual effect of KCl. The variations in the screening effect of the negative charges of the CF1 surface by K+ ions can indeed explain the changes in the anchorage of these 3 basic amino acids with concomitant variation in ATPase activity. Human thioredoxin must be 10 times more concentrated than Escherichia coli or spinach chloroplast thioredoxin to exhibit the same activation effect on the ATPase. This fact was related to the properties of a sequence equivalent to the part from amino acid 59 to 72 in Escherichia coli thioredoxin. This part which joins the two lobes of the thioredoxin is more hydrophilic and more negatively charged in human thioredoxin than in Escherichia coli or spinach chloroplast thioredoxin. Although ATPase activation was obtained at a very low concentration of the reduced spinach chloroplast thioredoxin, the thioredoxin formed only a loose complex with CF1.
authors: Service de Biophysique, CEN Saclay, Gif-sur-Yvette, France.
authors: J M
authors: JM
authors: Galmiche
authors: G
authors: G
authors: Girault
authors: G
authors: G
authors: Berger
authors: J P
authors: JP
authors: Jacquot
authors: M
authors: M
authors: Miginiac-Maslow
authors: E
authors: E
authors: Wollman
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000494:pharmacology*; D002736:Chloroplasts / Q000187:drug effects / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology*; D011189:Potassium Chloride / Q000494:pharmacology; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D013379:Substrate Specificity; D013879:Thioredoxins / Q000494:pharmacology*
chemical_list: D001426:Bacterial Proteins; D013879:Thioredoxins; D011189:Potassium Chloride; D006180:Proton-Translocating ATPases