title: Chloroplast fructose-1,6-bisphosphatase: modification of non-covalent interactions promote the activation by chimeric Escherichia coli thioredoxins.
abstract: Although all thioredoxins contain a highly conserved amino acid sequence responsible for thiol/disulfide exchanges, only chloroplast thioredoxin-f is effective in the reductive stimulation of chloroplast fructose-1,6-bisphosphatase. We set out to determine whether Escherichia coli thioredoxin becomes functional when selected modulators alter the conformation of the target enzyme. Wild type and chimeric Escherichia coli thioredoxins match the chloroplast counterpart when the activation of chloroplast fructose 1,6-biphosphatase is performed in the presence of fructose 1,6-bisphosphate, Ca2+, and either trichloroacetate or 2-propanol. These modulators of enzyme activity do change the conformation of chloroplast fructose-1,6-bisphosphatase whereas bacterial thioredoxins remain unaltered. Given that fructose 1,6-bisphosphate, Ca2+, and non-physiological perturbants modify non-covalent interactions of the protein but do not participate in redox reactions, these results strongly suggest that the conformation of the target enzyme regulates the rate of thiol/disulfide exchanges catalyzed by protein disulfide oxidoreductases.
authors.affiliation: Instituto de Investigaciones Bioquímicas (Fundación Campomar, I.I.B.-F.C.E.N.-U.B.A., IIBBA-CONICET), Buenos Aires, Argentina.
authors.forename: S
authors.initials: S
authors.lastname: Mora-García
authors.forename: M A
authors.initials: MA
authors.lastname: Ballícora
authors.forename: R A
authors.initials: RA
authors.lastname: Wolosiuk
mesh_terms: D000595:Amino Acid Sequence; D002736:Chloroplasts / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects*; D004926:Escherichia coli / Q000737:chemistry; D006597:Fructose-Bisphosphatase / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins; D013879:Thioredoxins / Q000737:chemistry / Q000494:pharmacology*; D014238:Trichloroacetic Acid / Q000494:pharmacology
chemical_list: D011993:Recombinant Fusion Proteins; D013879:Thioredoxins; D014238:Trichloroacetic Acid; D006597:Fructose-Bisphosphatase