Purification and properties of glutathione reductase from the cyanobacterium Anabaena sp. strain 7119.
An NADPH-glutathione reductase (EC 1.6.4.2) has been purified 6,000-fold to electrophoretic homogeneity from the filamentous cyanobacterium Anabaena sp. strain 7119. The purified enzyme exhibits a specific activity of 249 U/mg and is characterized by being a dimeric flavin adenine dinucleotide-containing protein with a ratio of absorbance at 280 nm to absorbance at 462 nm of 5.8, a native molecular weight of 104,000, a Stokes radius of 4.13 nm, and a pI of 4.02. The enzyme activity is inhibited by sulfhydryl reagents and heavy-metal ions, especially in the presence of NADPH, with oxidized glutathione behaving as a protective agent. As is the case with the same enzyme from other sources, the kinetic data are consistent with a branched mechanism. Nevertheless, the cyanobacterial enzyme presents three distinctive features with respect to that isolated from non-photosynthetic organisms: (i) absolute specificity for NADPH, (ii) an alkaline optimum pH value of ca. 9.0, and (iii) strong acidic character of the protein, as estimated by column chromatofocusing. The kinetic parameters are very similar to those found for the chloroplast enzyme, but the molecular weight is lower, being comparable to that of non-photosynthetic microorganisms. A protective function, analogous to that assigned to the chloroplast enzyme, is suggested.
D000596:Amino Acids / Q000032:analysis; D000458:Cyanobacteria / Q000201:enzymology*; D005182:Flavin-Adenine Dinucleotide / Q000032:analysis; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D019803:Glutathione Disulfide; D005980:Glutathione Reductase / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism / Q000494:pharmacology; D010084:Oxidation-Reduction; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology
D000596:Amino Acids; D013439:Sulfhydryl Reagents; D005182:Flavin-Adenine Dinucleotide; D009249:NADP; D005980:Glutathione Reductase; D005978:Glutathione; D015032:Zinc; D019803:Glutathione Disulfide