The plastocyanin binding domain of photosystem I.
The molecular recognition between plastocyanin and photosystem I was studied. Photosystem I and plastocyanin can be cross-linked to an active electron transfer complex. Immunoblots and mass spectrometric analysis of proteolytic peptides indicate that the two negative patches conserved in plant plastocyanins are cross-linked with lysine residues of a domain near the N-terminus of the PsaF subunit of photosystem I. Conversion of these negative to uncharged patches of plastocyanin by site-directed mutation D42N/E43Q/D44N/E45Q and E59Q/E60Q/D61N respectively, reveals the first patch to be essential for the electrostatic interaction in the electron transfer complex with photosystem I and the second one to lower the redox potential. The domain in PsaF, not found in cyanobacteria, is predicted to fold into two amphipathic alpha-helices. The interacting N-terminal helix lines up six lysines on one side which may guide a fast one-dimensional diffusion of plastocyanin and provide the electrostatic attraction at the attachment site, in addition to the hydrophobic interaction in the area where the electron is transferred to P700 in the reaction center of photosystem I. This two-step interaction is likely to increase the electron transfer rate by more than two orders of magnitude in plants as compared with cyanobacteria. Our data resolve the controversy about the function of PsaF.
Hippler
M
M
Biochemie der Pflanzen, Institut für Biologie II, Albert-Ludwigs-Universität, Freiburg, Germany.
Reichert
J
J
Sutter
M
M
Zak
E
E
Altschmied
L
L
Schröer
U
U
Herrmann
R G
RG
Haehnel
W
W
D000595:Amino Acid Sequence; D001665:Binding Sites; D003432:Cross-Linking Reagents; D000458:Cyanobacteria / Q000378:metabolism*; D003580:Cytochromes / Q000302:isolation & purification / Q000378:metabolism; D045348:Cytochromes f; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000378:metabolism*; D045331:Photosystem I Protein Complex; D010944:Plants / Q000378:metabolism; D010970:Plastocyanin / Q000378:metabolism*; D017354:Point Mutation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D018724:Spinacia oleracea / Q000378:metabolism*
D003432:Cross-Linking Reagents; D003580:Cytochromes; D010446:Peptide Fragments; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D011994:Recombinant Proteins; D010970:Plastocyanin; D045348:Cytochromes f