title: An examination of how structural changes can affect the rate of electron transfer in a mutated bacterial photoreaction centre.
abstract: A series of reaction centres bearing mutations at the (Phe) M197 position were constructed in the photosynthetic bacterium Rhodobacter sphaeroides. This residue is adjacent to the pair of bacteriochlorophyll molecules (P(L) and P(M)) that is the primary donor of electrons (P) in photosynthetic light-energy transduction. All of the mutations affected the optical and electrochemical properties of the P bacteriochlorophylls. A mutant reaction centre with the change Phe M197 to Arg (FM197R) was crystallized, and a structural model constructed at 2.3 A (1 A=0.1 nm) resolution. The mutation resulted in a change in the structure of the protein at the interface region between the P bacteriochlorophylls and the monomeric bacteriochlorophyll that is the first electron acceptor (B(L)). The new Arg residue at the M197 position undergoes a significant reorientation, creating a cavity at the interface region between P and B(L). The acetyl carbonyl substituent group of the P(M) bacteriochlorophyll undergoes an out-of-plane rotation, which decreases the edge-to-edge distance between the macrocycles of P(M) and B(L). In addition, two new buried water molecules partially filled the cavity that is created by the reorientation of the Arg residue. These waters are in a suitable position to connect the macrocycles of P and B(L) via three hydrogen bonds. Transient absorption measurements show that, despite an inferred decrease in the driving force for primary electron transfer in the FM197R reaction centre, there is little effect on the overall rate of the primary reaction in the bulk of the reaction-centre population. Examination of the X-ray crystal structure reveals a number of small changes in the structure of the reaction centre in the interface region between the P and B(L) bacteriochlorophylls that could account for this faster-than-predicted rate of primary electron transfer.
authors.affiliation: Department of Molecular Biology and Biotechnology, University of Sheffield, Western Bank, Sheffield S10 2UH, U.K.
authors.forename: J P
authors.initials: JP
authors.lastname: Ridge
authors.forename: P K
authors.initials: PK
authors.lastname: Fyfe
authors.forename: K E
authors.initials: KE
authors.lastname: McAuley
authors.forename: M E
authors.initials: ME
authors.lastname: van Brederode
authors.forename: B
authors.initials: B
authors.lastname: Robert
authors.forename: R
authors.initials: R
authors.lastname: van Grondelle
authors.forename: N W
authors.initials: NW
authors.lastname: Isaacs
authors.forename: R J
authors.initials: RJ
authors.lastname: Cogdell
authors.forename: M R
authors.initials: MR
authors.lastname: Jones
mesh_terms: D018360:Crystallography, X-Ray; D004579:Electron Transport; D006860:Hydrogen Bonding; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes; D016297:Mutagenesis, Site-Directed; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins