Crystal structure of the wild-type and D30A mutant thioredoxin h of Chlamydomonas reinhardtii and implications for the catalytic mechanism
Autor: | V, Menchise, C, Corbier, C, Didierjean, M, Saviano, E, Benedetti, J P, Jacquot, A, Aubry |
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Rok vydání: | 2001 |
Předmět: |
Models
Molecular Aspartic Acid Binding Sites Sequence Homology Amino Acid Protein Conformation Molecular Sequence Data Thioredoxin h Crystallography X-Ray Thioredoxins Mutation Animals Humans Computer Simulation Amino Acid Sequence Protons Oxidation-Reduction Chlamydomonas reinhardtii Conserved Sequence Research Article |
Zdroj: | The Biochemical journal. 359(Pt 1) |
ISSN: | 0264-6021 |
Popis: | Thioredoxins are ubiquitous proteins which catalyse the reduction of disulphide bridges on target proteins. The catalytic mechanism proceeds via a mixed disulphide intermediate whose breakdown should be enhanced by the involvement of a conserved buried residue, Asp-30, as a base catalyst towards residue Cys-39. We report here the crystal structure of wild-type and D30A mutant thioredoxin h from Chlamydomonas reinhardtii, which constitutes the first crystal structure of a cytosolic thioredoxin isolated from a eukaryotic plant organism. The role of residue Asp-30 in catalysis has been revisited since the distance between the carboxylate OD1 of Asp-30 and the sulphur SG of Cys-39 is too great to support the hypothesis of direct proton transfer. A careful analysis of all available crystal structures reveals that the relative positioning of residues Asp-30 and Cys-39 as well as hydrophobic contacts in the vicinity of residue Asp-30 do not allow a conformational change sufficient to bring the two residues close enough for a direct proton transfer. This suggests that protonation/deprotonation of Cys-39 should be mediated by a water molecule. Molecular-dynamics simulations, carried out either in vacuo or in water, as well as proton-inventory experiments, support this hypothesis. The results are discussed with respect to biochemical and structural data. |
Databáze: | OpenAIRE |
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