Sulfolobus solfataricus protein disulphide oxidoreductase: insight into the roles of its redox sites(444 views) Limauro D, Saviano M, Galdi I, Rossi M, Bartolucci S, Pedone E
Keywords: Protein Disulphide Oxidoreductase, Redox Sites, Sulfolobus Solfataricus, Thioredoxin Fold, Thioredoxin System, Accessible Surfaces, Biochemical Analysis, Circular Dichroism Datums, Computational Analyses, Cys Residues, Cysteine Residues, Disulphide Bridges, Functional Features, Functional Properties, Peroxiredoxin, Pk Values, Reduced Forms, Reductase Activities, Side Chains, Single Mutants, Solvent Exposures, Thioredoxin Reductases, Three-Dimensional Models, Enzyme Activity, Optical Properties, Three Dimensional, Protein Folding, Bacterial Enzyme, Protein Disulfide Oxidoreductase, Unclassified Drug, Article, Nonhuman, Priority Journal, Protein Function, Protein Stability, Amino Acid Sequence, Catalytic Domain, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Conformation, Protein Disulfide Reductase (glutathione), Protein Engineering, Sequence Alignment,
Affiliations: *** IBB - CNR ***
Dip. Biologia Strutturale e Funzionale, University of Naples Federico II, Complesso Universitario Monte S. Angelo, via Cinthia, 80126 Naples, Italy
Istituto di Biostrutture e Bioimmagini, C.N.R., Via Mezzocannone 16, 80134 Naples, Italy
References: Not available.
Sulfolobus solfataricus protein disulphide oxidoreductase: insight into the roles of its redox sites
Sulfolobus solfataricus protein disulphide oxidoreductase (SsPDO) contains three disulphide bridges linking residues C(41)XXC(44), C(155)XXC(158), C(173)XXXXC(178). To get information on the role played by these cross-links in determining the structural and functional properties of the protein, we performed site-directed mutagenesis on Cys residues and investigated the changes in folding, stability and functional features of the mutants and analysed the results with computational analysis. The reductase activity of SsPDO and its mutants was evaluated by insulin and thioredoxin reductase assays also coupled with peroxiredoxin Bcp1 of S. solfataricus. The three-dimensional model of SsPDO was constructed and correlated with circular dichroism data and functional results. Biochemical analysis indicated a key function for the redox site constituted by Cys155 and Cys158. To discriminate between the role of the two cysteine residues, each cysteine was mutagenised and the behaviour of the single mutants was investigated elucidating the basis of the electron-shuffling mechanism for SsPDO. Finally, cysteine pK values were calculated and the accessible surface for the cysteine side chains in the reduced form was measured, showing higher reactivity and solvent exposure for Cys155.
Sulfolobus solfataricus protein disulphide oxidoreductase: insight into the roles of its redox sites
No results.
Sulfolobus solfataricus protein disulphide oxidoreductase: insight into the roles of its redox sites