Purification and characterization of thioredoxin from the N2-fixing cyanobacterium Anabaena cylindrica.
Thioredoxin has been purified to homogeneity from the cyanobacterium Anabaena cylindrica. The protein consists of a single polypeptide chain with a relative molecular mass of about 11 680 which has two cysteine residues (residues 31 and 34) in the sequence-Cys-Gly-Pro-Cys- and an isoelectric point at pH 4.55. The N-terminal amino acid sequence of 39 residues shows distinct homologies with the sequences of Escherichia coli and Corynebacterium nephridii thioredoxins. Anti-(A. cylindrica thioredoxin) antiserum was used to quantify the thioredoxin which constituted about 0.22% of the soluble protein in cell-free extracts of N2-fixing, NO3- -grown or NH4+-grown A. cylindrica. Activation of fructose-1,6-bisphosphatase of A. cylindrica, activation of glutamine synthetase and NADP+-dependent malate dehydrogenase of the green alga Scenedesmus obliquus but not of A. cylindrica, and deactivation of glucose-6-P dehydrogenase of the cyanobacterium Anabaena variabilis were all achieved using the same thioredoxin species. No other thioredoxin species were detected in extracts of A. cylindrica when examined for the activation of these enzymes.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002621:Chemistry; D000458:Cyanobacteria / Q000032:analysis* / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods; D006597:Fructose-Bisphosphatase / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D007122:Immunoelectrophoresis / Q000379:methods; D008291:Malate Dehydrogenase / Q000378:metabolism; D009586:Nitrogen Fixation; D013879:Thioredoxins / Q000302:isolation & purification* / Q000378:metabolism
D000596:Amino Acids; D001426:Bacterial Proteins; D013879:Thioredoxins; D008291:Malate Dehydrogenase; C031917:malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+); D005954:Glucosephosphate Dehydrogenase; D006597:Fructose-Bisphosphatase; D005974:Glutamate-Ammonia Ligase