The unique proline of the Prochlorothrix hollandica plastocyanin hydrophobic patch impairs electron transfer to photosystem I.
A number of surface residues of plastocyanin from Prochlorothrix hollandica have been modified by site-directed mutagenesis. Changes have been made in amino acids located in the amino-terminal hydrophobic patch of the copper protein, which presents a variant structure as compared with other plastocyanins. The single mutants Y12G, Y12F, Y12W, P14L, and double mutant Y12G/P14L have been produced. Their reactivity toward photosystem I has been analyzed by laser flash absorption spectroscopy. Plots of the observed rate constant with all mutants versus plastocyanin concentration show a saturation profile similar to that with wild-type plastocyanin, thus suggesting the formation of a plastocyanin-photosystem I transient complex. The mutations do not induce relevant changes in the equilibrium constant for complex formation but induce significant variations in the electron transfer rate constant, mainly with the two mutants at proline 14. Additionally, molecular dynamics calculations indicate that mutations at position 14 yield small changes in the geometry of the copper center. The comparative kinetic analysis of the reactivity of plastocyanin mutants toward photosystem I from different organisms (plants and cyanobacteria) reveals that reversion of the unique proline of Prochlorothrix plastocyanin to the conserved leucine of all other plastocyanins at this position enhances the reactivity of the Prochlorothrix protein.
Navarro
J A
JA
Instituto de Bioquimica Vegetal y Fotosintesis, Centro de Investigaciones Cientificas Isla de la Cartuja, Universidad de Sevilla, Spain.
Myshkin
E
E
De la Rosa
M A
MA
Bullerjahn
G S
GS
Hervás
M
M
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004579:Electron Transport; D007700:Kinetics; D020409:Molecular Motor Proteins; D009154:Mutation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D045331:Photosystem I Protein Complex; D010970:Plastocyanin / Q000235:genetics / Q000378:metabolism*; D019692:Prochlorothrix / Q000235:genetics / Q000378:metabolism*; D011392:Proline / Q000378:metabolism*
D001426:Bacterial Proteins; D020409:Molecular Motor Proteins; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; C070733:photosystem I, core protein, Synechococcus; D010970:Plastocyanin; D011392:Proline