title: Enzyme regulation in C4 photosynthesis: purification, properties, and activities of thioredoxins from C4 and C3 plants.
abstract: Procedures are described for the purification to homogeneity of chloroplast thioredoxins f and m from leaves of corn (Zea mays, a C4 plant) and spinach (Spinacea oleracea, a C3 plant). The C3 and C4f thioredoxins were similar immunologically and biochemically, but differed in certain of their physiochemical properties. The f thioredoxins from the two species were capable of activating both NADP-malate dehydrogenase (EC 1.1.1.37) and fructose-1,6-bisphosphatase (EC 3.1.3.11) when tested in standard thioredoxin assays. Relative to its spinach counterpart, corn thioredoxin f showed a greater molecular mass (15.0-16.0 kDa vs 10.5 kDa), lower isoelectric point (ca. 5.2 vs 6.0), and lower ability to form a stable noncovalent complex with its target fructose bisphosphatase enzyme. The C3 and C4 m thioredoxins were similar in their specificity (ability to activate NADP-malate dehydrogenase, and not fructose-1,6-bisphosphatase) and isoelectric points (ca. 4.8), but differed slightly in molecular mass (13.0 kDa for spinach vs 13.5 kDa for corn) and substantially in their immunological properties. Results obtained in conjunction with these studies demonstrated that the thioredoxin m-linked activation of NADP-malate dehydrogenase in selectively enhanced by the presence of halide ions (e.g., chloride) and by an organic solvent (e.g., 2-propanol). The results suggest that in vivo NADP-malate dehydrogenase interacts with thylakoid membranes and is regulated to a greater extent by thioredoxin m than thioredoxin f.
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification*; D054479:Chloroplast Thioredoxins; D002736:Chloroplasts / Q000378:metabolism; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006597:Fructose-Bisphosphatase / Q000378:metabolism; D007120:Immunochemistry; D007526:Isoelectric Point; D008291:Malate Dehydrogenase / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D010788:Photosynthesis; D010940:Plant Proteins / Q000302:isolation & purification* / Q000502:physiology; D010944:Plants / Q000378:metabolism; D011485:Protein Binding; D012492:Salts / Q000494:pharmacology; D012997:Solvents / Q000494:pharmacology; D013879:Thioredoxins / Q000302:isolation & purification* / Q000502:physiology; D003313:Zea mays / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D054479:Chloroplast Thioredoxins; D010940:Plant Proteins; D012492:Salts; D012997:Solvents; D013879:Thioredoxins; D009249:NADP; D008291:Malate Dehydrogenase; D006597:Fructose-Bisphosphatase