A single mutation in the heme 4 environment of Desulfovibrio desulfuricans Norway cytochrome c3 (Mr 26,000) greatly affects the molecule reactivity.
The gene encoding Desulfovibrio desulfuricans Norway cytochrome c3 (Mr 26,000), a dimeric octaheme cytochrome belonging to the polyheme cytochrome c3 superfamily, has been cloned and successfully expressed in another sulfate reducing bacteria, D. desulfuricans G201. The gene, named cycD, is monocistronic and encodes a cytochrome precursor of 135 amino acids with an extension at the NH2 terminus of 24 amino acids. This extension acts as a signal sequence which allows export across the cytoplasmic membrane into the periplasmic space. Tyrosine 73, which is in a close contact with the histidine sixth axial ligand to the heme 4 iron atom, has been replaced by a glutamate residue using site-directed mutagenesis. The cytochrome mutant when expressed in D. desulfuricans G201, is correctly folded and matured. A global increase of the oxidoreduction potentials of about 50 mV is measured for the Y73E cytochrome. The mutation also has a strong influence on the interaction of the cytochrome with its redox partner, the hydrogenase. This suggests, like the tetraheme cytochrome c3 (Mr 13, 000), heme 4 is the interactive heme in the cytochrome-hydrogenase complex and that alteration of the heme 4 environment can greatly affect the electron transfer reaction with its redox partner.
Aubert
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Unité de Bioénergétique et Ingéniérie des Protéines, IFR1-Centre National de la Recherche Scientifique, 13402 Marseille Cedex 20, France.
Leroy
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Bruschi
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Wall
J D
JD
Dolla
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A
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial; D003901:Desulfovibrio / Q000201:enzymology / Q000235:genetics*; D006418:Heme / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed
D003574:Cytochrome c Group; D004269:DNA, Bacterial; D006418:Heme; C017165:cytochrome c(3)