Overexpression of Desulfovibrio vulgaris Hildenborough cytochrome c553 in Desulfovibrio desulfuricans G200. Evidence of conformational heterogeneity in the oxidized protein by NMR.
Plasmid pRC41, containing the cyf gene encoding cytochrome c533 from Desulfovibrio vulgaris Hildenborough, was transferred by conjugation from Escherichia coli to Desulfovibrio desulfuricans G200. The structural properties of the purified protein were studied by one-dimensional and two-dimensional NMR. A heterogeneity in the folding of the cytochrome isolated from D. vulgaris Hildenborough and from D. desulfuricans G200 was observed for the oxidized from. Temperature, pH and salt-dependence studies indicated that the heterogeneity does not result from an intermediate in the protein unfolding process, but derives from two conformations which are not in dynamic equilibrium.
Blanchard
L
L
Laboratoire de Chimie Bactérienne, CNRS, Marseille, France.
Marion
D
D
Pollock
B
B
Voordouw
G
G
Wall
J
J
Bruschi
M
M
Guerlesquin
F
F
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D004269:DNA, Bacterial; D003901:Desulfovibrio / Q000235:genetics*; D016969:Desulfovibrio vulgaris / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012492:Salts; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D003574:Cytochrome c Group; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D012492:Salts; C083951:cytochrome c553; C022302:cytochrome C-552