title: Tuning of the optical and electrochemical properties of the primary donor bacteriochlorophylls in the reaction centre from Rhodobacter sphaeroides: spectroscopy and structure.
abstract: A series of mutations have been introduced at residue 168 of the L-subunit of the reaction centre from Rhodobacter sphaeroides. In the wild-type reaction centre, residue His L168 donates a strong hydrogen bond to the acetyl carbonyl group of one of the pair of bacteriochlorophylls (BChl) that constitutes the primary donor of electrons. Mutation of His L168 to Phe or Leu causes a large decrease in the mid-point redox potential of the primary electron donor, consistent with removal of this strong hydrogen bond. Mutations to Lys, Asp and Arg cause smaller decreases in redox potential, indicative of the presence of weak hydrogen bond and/or an electrostatic effect of the polar residue. A spectroscopic analysis of the mutant complexes suggests that replacement of the wild-type His residue causes a decrease in the strength of the coupling between the two primary donor bacteriochlorophylls. The X-ray crystal structure of the mutant in which His L168 has been replaced by Phe (HL168F) was determined to a resolution of 2.5 A, and the structural model of the HL168F mutant was compared with that of the wild-type complex. The mutation causes a shift in the position of the primary donor bacteriochlorophyll that is adjacent to residue L168, and also affects the conformation of the acetyl carbonyl group of this bacteriochlorophyll. This conformational change constitutes an approximately 27 degrees through-plane rotation, rather than the large into-plane rotation that has been widely discussed in the context of the HL168F mutation. The possible structural basis of the altered spectroscopic properties of the HL168F mutant reaction centre is discussed, as is the relevance of the X-ray crystal structure of the HL168F mutant to the possible structures of the remaining mutant complexes.
authors.affiliation: Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, Sheffield S10 2UH, UK.
authors.forename: Diane
authors.initials: D
authors.lastname: Spiedel
authors.forename: Aleksander W
authors.initials: AW
authors.lastname: Roszak
authors.forename: Kimberley
authors.initials: K
authors.lastname: McKendrick
authors.forename: Katherine E
authors.initials: KE
authors.lastname: McAuley
authors.forename: Paul K
authors.initials: PK
authors.lastname: Fyfe
authors.forename: Eliane
authors.initials: E
authors.lastname: Nabedryk
authors.forename: Jacques
authors.initials: J
authors.lastname: Breton
authors.forename: Bruno
authors.initials: B
authors.lastname: Robert
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Cogdell
authors.forename: Neil W
authors.initials: NW
authors.lastname: Isaacs
authors.forename: Michael R
authors.initials: MR
authors.lastname: Jones
mesh_terms: D001429:Bacteriochlorophylls / Q000737:chemistry*; D018360:Crystallography, X-Ray; D004563:Electrochemistry; D045342:Light-Harvesting Protein Complexes; D008958:Models, Molecular; D009154:Mutation; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D012242:Rhodobacter sphaeroides / Q000737:chemistry*; D013054:Spectrophotometry, Atomic; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D001429:Bacteriochlorophylls; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins