Dynamic interaction of plastocyanin with the cytochrome bf complex.
The interaction between plastocyanin and the intact cytochrome bf complex, both from spinach, has been studied by stopped-flow kinetics with mutant plastocyanin to elucidate the site of electron transfer and the docking regions of the molecule. Mutation of Tyr-83 to Arg or Leu provides no evidence for a second electron transfer path via Tyr-83 of plastocyanin, which has been proposed to be the site of electron transfer from cytochrome f. The data found with mutations of acidic residues indicate that both conserved negative patches are essential for the binding of plastocyanin to the intact cytochrome bf complex. Replacing Ala-90 and Gly-10 at the flat hydrophobic surface of plastocyanin by larger residues slowed down and accelerated, respectively, the rate of electron transfer as compared with wild-type plastocyanin. These opposing effects reveal that the hydrophobic region around the electron transfer site at His-87 is divided up into two regions, of which only that with Ala-90 contributes to the attachment to the cytochrome bf complex. These binding sites of plastocyanin are substantially different from those interacting with photosystem I. It appears that each of the two binding regions of plastocyanin is split into halves, which are used in different combinations in the molecular recognition at the two membrane complexes.
Illerhaus
J
J
Institut für Biologie II/Biochemie, Albert-Ludwigs-Universität Freiburg, Schänzlestrasse 1, D-79104 Freiburg, Germany.
Altschmied
L
L
Reichert
J
J
Zak
E
E
Herrmann
R G
RG
Haehnel
W
W
D019943:Amino Acid Substitution; D001120:Arginine; D003573:Cytochrome b Group / Q000737:chemistry* / Q000378:metabolism*; D045346:Cytochrome b6f Complex; D007700:Kinetics; D007930:Leucine; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010084:Oxidation-Reduction; D010970:Plastocyanin / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D018724:Spinacia oleracea / Q000378:metabolism; D014443:Tyrosine
D003573:Cytochrome b Group; D011994:Recombinant Proteins; D014443:Tyrosine; D010970:Plastocyanin; D045346:Cytochrome b6f Complex; D001120:Arginine; D007930:Leucine