Thioredoxin-linked processes in cyanobacteria are as numerous as in chloroplasts, but targets are different.
Light-dependent regulation of a growing number of chloroplast enzymatic activities has been found to occur through the reversible reduction of intra- or intermolecular disulphides by thioredoxins. In cyanobacteria, despite their similarity to chloroplasts, no proteins have hitherto been shown to interact with thioredoxins, and the role of the cyanobacterial ferredoxin/thioredoxin system has remained obscure. By using an immobilized cysteine 35-to-serine site-directed mutant of the Synechocystis sp. PCC 6803 thioredoxin TrxA as bait, we screened the Synechocystis cytosolic and peripheral membrane protein complements for proteins interacting with TrxA. The covalent bond between the isolated target proteins and mutated TrxA was confirmed by nonreducing/reducing two-dimensional SDS/PAGE. Thus, we have identified 18 cytosolic proteins and 8 membrane-associated proteins as candidate thioredoxin substrates. Twenty of these proteins have not previously been associated with thioredoxin-mediated regulation. Phosphoglucomutase, one of the previously uncharacterized thioredoxin-linked enzymes, has not earlier been considered a target for metabolic control through disulphide reduction. In this article, we show that phosphoglucomutase is inhibited under oxidizing conditions and activated by DTT and reduced wild-type TrxA in vitro. The results imply that thioredoxin-mediated redox regulation is as extensive in cyanobacteria as in chloroplasts but that the subjects of regulation are largely different.
Lindahl
Marika
M
Instituto de Bioquímica Vegetal y Fotosíntesis, Consejo Superior de Investigaciones Científicas, Universidad de Sevilla, Centro de Investigaciones Científicas Isla de la Cartuja, Avenida Americo Vespucio s/n, E-410 92 Sevilla, Spain. lindahl@ibvf.csic.es
Florencio
Francisco J
FJ
D001426:Bacterial Proteins / Q000378:metabolism; D002736:Chloroplasts / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D000458:Cyanobacteria / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D005288:Ferredoxins / Q000378:metabolism*; D010084:Oxidation-Reduction; D010733:Phosphoglucomutase / Q000378:metabolism; D013879:Thioredoxins / Q000378:metabolism*
D001426:Bacterial Proteins; D005288:Ferredoxins; D013879:Thioredoxins; D010733:Phosphoglucomutase; D004229:Dithiothreitol