A single mutation in the heme 4 environment of Desulfovibrio desulfuricans Norway cytochrome c3 (Mr 26,000) greatly affects the molecule reactivity.

The Journal of Biological Chemistry
C AubertA Dolla

Abstract

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 Tyrosine 73, which is in a close contact with the histidine sixth axial ligand to the heme 4 iron atom, has been replaced by a glutamate residue using site-directed mutagenesis.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 tran...Continue Reading

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Citations

Jan 9, 2004·Applied and Environmental Microbiology·Dwayne A EliasLee R Krumholz
Oct 18, 2005·Journal of Molecular Biology·Laetitia PieulleE Claude Hatchikian
May 11, 2004·Protein Science : a Publication of the Protein Society·P Raj PokkuluriMarianne Schiffer
Jan 24, 1998·Biochemical and Biophysical Research Communications·C AubertA Dolla
Dec 22, 1999·Biochimica Et Biophysica Acta·C AubertM T Giudici-Orticoni

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