Identification of the site of interaction between cytochrome c3 and ferredoxin using peptide mapping of the cross-linked complex.
Structural studies carried out on a cross-linked complex between cytochrome c3 and ferredoxin I, both isolated from Desulfovibrio desulfuricans Norway, allowed the identification of the site of interaction between the two redox proteins. Staphylococcus aureus proteinase and chymotrypsin digestions led to characterization of peptides containing both cytochrome c3 and ferredoxin sequences. The cytochrome c3 sequences involved in the three isolated cross-linked peptides contained several lysine residues localized around the heme 4 crevice. This analysis stressed the peculiar role of lysines 100, 101, 103, 104 and 113, which could be considered as major cross-link sites, as opposed to the lysines 75, 79 and 82, which could be considered as minor cross-link sites. One cross-linked peptide, containing two ferredoxin sequences joined to one cytochrome c3 sequence, had been isolated, suggesting the possibility of more than one cross-link per covalent complex. All these results led to the identification of heme 4 of cytochrome c3 as the site of interaction for the ferredoxin I. This study confirms the proposal that could be deduced from the hypothetical structure of the complex built by computer graphics modelling (Cambillau, C., Frey, M., Mosse, J., Guerlesquin, F. and Bruschi, M. (1988) Proteins: struct., funct. genet. 4, 63-70).
Dolla
A
A
Laboratoire de Chimie Bactérienne du CNRS, Marseille, France.
Leroy
G
G
Guerlesquin
F
F
Bruschi
M
M
D001665:Binding Sites; D002918:Chymotrypsin / Q000737:chemistry; D003432:Cross-Linking Reagents; D003574:Cytochrome c Group / Q000378:metabolism*; D003901:Desulfovibrio / Q000201:enzymology / Q000378:metabolism; D010450:Endopeptidases / Q000737:chemistry; D005288:Ferredoxins / Q000378:metabolism*; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010449:Peptide Mapping; D011487:Protein Conformation; D013211:Staphylococcus aureus / Q000201:enzymology
D003432:Cross-Linking Reagents; D003574:Cytochrome c Group; D005288:Ferredoxins; C017165:cytochrome c(3); D010450:Endopeptidases; D002918:Chymotrypsin