Site-directed mutagenesis of tetraheme cytochrome c3. Modification of oxidoreduction potentials after heme axial ligand replacement
| Autor: | Richard Haser, Françoise Payan, J. Haladjian, Pierre Bianco, Mirjam Czjzek, Isabelle Mus-Veteau, J D Wall, Alain Dolla, Françoise Guerlesquin, B J Rapp-Giles |
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| Přispěvatelé: | Deleage, Gilbert |
| Rok vydání: | 1992 |
| Předmět: |
Models
Molecular Hemeprotein Magnetic Resonance Spectroscopy Cytochrome Stereochemistry Protein Conformation Genetic Vectors Molecular Sequence Data Restriction Mapping Cytochrome c Group Heme Ligands Biochemistry chemistry.chemical_compound [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology Amino Acid Sequence Desulfovibrio vulgaris Site-directed mutagenesis Molecular Biology Binding Sites biology Base Sequence Cytochrome b Cytochrome c Mutagenesis Cell Biology biology.organism_classification chemistry Oligodeoxyribonucleotides biology.protein Mutagenesis Site-Directed |
| Zdroj: | The Journal of biological chemistry. 267(24) |
| ISSN: | 0021-9258 |
| Popis: | The nature of the axial ligands of a heme group is an important factor in maintaining the oxidation-reduction potential of a c-type cytochrome. Cytochrome c3 from Desulfovibrio vulgaris Hildenborough contains four bis-histidinyl coordinated hemes with low oxidation-reduction potentials. Site-directed mutagenesis was used to generate a mutant in which histidine 70, the sixth axial ligand of heme 4, has been replaced by a methionine. The mutant protein was expressed in Desulfovibrio desulfuricans G200 at a level similar to the wild type cytochrome. A model for the three-dimensional structure of D. vulgaris Hildenborough cytochrome c3 was generated on the basis of the crystal structure of D. vulgaris Miyazaki cytochrome c3 in order to investigate the effects of the H70M mutation. The model, together with NMR data, suggested that methionine 70 has effectively replaced histidine 70 as the sixth axial ligand of heme 4 without significant alteration of the structure. A large increase of at least 200 mV of one of the four oxidation-reduction potentials was observed by electrochemistry and is interpreted in terms of structure/potential relationships.The nature of the axial ligands of a heme group is an important factor in maintaining the oxidation-reduction potential of a c-type cytochrome. Cytochrome c3 from Desulfovibrio vulgaris Hildenborough contains four bis-histidinyl coordinated hemes with low oxidation-reduction potentials. Site-directed mutagenesis was used to generate a mutant in which histidine 70, the sixth axial ligand of heme 4, has been replaced by a methionine. The mutant protein was expressed in Desulfovibrio desulfuricans G200 at a level similar to the wild type cytochrome. A model for the three-dimensional structure of D. vulgaris Hildenborough cytochrome c3 was generated on the basis of the crystal structure of D. vulgaris Miyazaki cytochrome c3 in order to investigate the effects of the H70M mutation. The model, together with NMR data, suggested that methionine 70 has effectively replaced histidine 70 as the sixth axial ligand of heme 4 without significant alteration of the structure. A large increase of at least 200 mV of one of the four oxidation-reduction potentials was observed by electrochemistry and is interpreted in terms of structure/potential relationships. |
| Databáze: | OpenAIRE |
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