title: Contributions of the protein environment to the midpoint potentials of the A1 phylloquinones and the Fx iron-sulfur cluster in photosystem I.
abstract: Electrostatic calculations have predicted that the partial negative charge associated with D575PsaB plays a significant role in modulating the midpoint potentials of the A1A and A1B phylloquinones in photosystem I. To test this prediction, the side chain of residue 575PsaB was changed from negatively charged (D) to neutral (A) and to positively charged (K). D566PsaB, which is located at a considerable distance from either A1A or A1B, and should affect primarily the midpoint potential of FX, was similarly changed. In the 575PsaB variants, the rate of electron transfer from A1A to FX is observed to decrease slightly according to the sequence D/A/K. In the 566PsaB variants, the opposite effect of a slight increase in the rate is observed according to the same sequence D/A/K. These results are consistent with the expectation that changing these residues will shift the midpoint potentials of nearby cofactors to more positive values and that the magnitude of this shift will depend on the distance between the cofactors and the residues being changed. Thus, the midpoint potentials of A1A and A1B should experience a larger shift than will FX in the 575PsaB variants, while FX should experience a larger shift than will either A1A or A1B in the 566PsaB variants. As a result, the driving energy for electron transfer from A1A and A1B to FX will be decreased in the former and increased in the latter. This rationalization of the changes in kinetics is compared with the results of electrostatic calculations. While the altered amino acids shift the midpoint potentials of A1A, A1B, and FX by different amounts, the difference in the shifts between A1A and FX or between A1B and FX is small so that the overall effect on the electron transfer rate between A1A and FX or between A1B and FX is predicted to be small. These conclusions are borne out by experiment.
authors.affiliation: Institut für Experimentalphysik, Freie Universität Berlin, Arnimallee 14, D-14195, Berlin, Germany.
authors.forename: Irina
authors.initials: I
authors.lastname: Karyagina
authors.forename: Yulia
authors.initials: Y
authors.lastname: Pushkar
authors.forename: Dietmar
authors.initials: D
authors.lastname: Stehlik
authors.forename: Art
authors.initials: A
authors.lastname: van der Est
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ishikita
authors.forename: Ernst-Walter
authors.initials: EW
authors.lastname: Knapp
authors.forename: Bharat
authors.initials: B
authors.lastname: Jagannathan
authors.forename: Rufat
authors.initials: R
authors.lastname: Agalarov
authors.forename: John H
authors.initials: JH
authors.lastname: Golbeck
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D004579:Electron Transport; D005288:Ferredoxins / Q000737:chemistry / Q000378:metabolism*; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D007506:Iron-Sulfur Proteins / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010084:Oxidation-Reduction; D045331:Photosystem I Protein Complex / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D011199:Potentiometry; D011487:Protein Conformation; D010837:Vitamin K 1 / Q000737:chemistry / Q000378:metabolism*
chemical_list: D005288:Ferredoxins; D007506:Iron-Sulfur Proteins; D045342:Light-Harvesting Protein Complexes; D045331:Photosystem I Protein Complex; C089909:photosystem I, psaB subunit; D010837:Vitamin K 1