title: Strong effects of an individual water molecule on the rate of light-driven charge separation in the Rhodobacter sphaeroides reaction center.
abstract: The role of a water molecule (water A) located between the primary electron donor (P) and first electron acceptor bacteriochlorophyll (B(A)) in the purple bacterial reaction center was investigated by mutation of glycine M203 to leucine (GM203L). The x-ray crystal structure of the GM203L reaction center shows that the new leucine residue packs in such a way that water A is sterically excluded from the complex, but the structure of the protein-cofactor system around the mutation site is largely undisturbed. The results of absorbance and resonance Raman spectroscopy were consistent with either the removal of a hydrogen bond interaction between water A and the keto carbonyl group of B(A) or a change in the local electrostatic environment of this carbonyl group. Similarities in the spectroscopic properties and x-ray crystal structures of reaction centers with leucine and aspartic acid mutations at the M203 position suggested that the effects of a glycine to aspartic acid substitution at the M203 position can also be explained by steric exclusion of water A. In the GM203L mutant, loss of water A was accompanied by an approximately 8-fold slowing of the rate of decay of the primary donor excited state, indicating that the presence of water A is important for optimization of the rate of primary electron transfer. Possible functions of this water molecule are discussed, including a switching role in which the redox potential of the B(A) acceptor is rapidly modulated in response to oxidation of the primary electron donor.
authors.affiliation: Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, Bristol, BS8 1TD, United Kingdom.
authors.forename: Jane A
authors.initials: JA
authors.lastname: Potter
authors.forename: Paul K
authors.initials: PK
authors.lastname: Fyfe
authors.forename: Dmitrij
authors.initials: D
authors.lastname: Frolov
authors.forename: Marion C
authors.initials: MC
authors.lastname: Wakeham
authors.forename: Rienk
authors.initials: R
authors.lastname: van Grondelle
authors.forename: Bruno
authors.initials: B
authors.lastname: Robert
authors.forename: Michael R
authors.initials: MR
authors.lastname: Jones
mesh_terms: D019943:Amino Acid Substitution; D001429:Bacteriochlorophylls / Q000737:chemistry*; D018360:Crystallography, X-Ray; D004579:Electron Transport / Q000235:genetics / Q000528:radiation effects; D006860:Hydrogen Bonding; D007700:Kinetics; D008027:Light; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000235:genetics; D012242:Rhodobacter sphaeroides / Q000737:chemistry*; D013057:Spectrum Analysis; D014867:Water / Q000737:chemistry*
chemical_list: D001429:Bacteriochlorophylls; D045322:Photosynthetic Reaction Center Complex Proteins; D014867:Water