title: Resonance Raman characterization of H(M200)L mutant reaction centers from Rhodobacter capsulatus. Effects of heterodimer formation on the structural and electronic properties of the cofactors.
abstract: Resonance Raman (RR) spectra are reported for photosynthetic reactions centers (RCs) from the H(M200)L mutant of Rhodobacter capsulatus. In this mutant, the histidine residue which ligates the M-side bacteriochlorophyll (BCh) of the special pair primary donor (P) of wild-type RCs is replaced by a noncoordinating leucine. This results in the formation of a heterodimer primary donor (D) in which a bacteriopheophytin (BPh) replaces the M-side BCh. The RR data for the H(M200)L mutant were acquired at a large number of excitation wavelengths which span the B, Qx, and Qy absorption bands of the various bacteriochlorin cofactors in the RC. For comparison, spectra were also acquired for wild-type RCs at the same excitation wavelengths. The RR data obtained for the mutant indicate that heterodimer formation induces a variety of changes in the structural and electronic properties of the cofactors in the RC. These perturbations extend beyond the primary donor and include one of the two accessory BChs. Collectively, the RR studies indicate the following: (1) The structure of the single BCh cofactor in D [DL(BCh)] is different from that of either of the two BChs in P. However, DL(BCh) is more similar to PL than to PM. The PM cofactor is conformationally more distorted than either PL or DL(BCh). (2) The structure of the BPh cofactor in D [DM(BPh)] is similar to that of the other two BPhs in the RC. However, the frequency of the C9-keto carbonyl mode of DM(BPh) is anomalously low (1678 cm-1), as is also the case for PM. The vibrational characteristics of the C9-keto carbonyl vibrations of DM(BPh)/PM versus DL(BCh)/PL are consistent the notion that dielectric effects govern the frequency of the mode and that the effective dielectric constant is different on the L- versus M-sides of the primary donor. (3) Heterodimer formation perturbs the structural and electronic properties of one of the two accessory BChs (most likely BChL) in the RC. These perturbations are manifested as upshifts in the ring skeletal-mode frequencies and a blue-shift in the Qx absorption band (from 600 to 580 nm). The fact that heterodimer formation perturbs one of the accessory BChs suggests that global structural rearrangements occur in the protein matrix when the ligand to a cofactor in the primary donor is removed. (4) For both the H(M200)L mutant and wild-type RCs, oxidation of the primary donor significantly affects the RR cross section of the carotenoid.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Chemistry, University of California, Riverside 92521-0403, USA.
authors.forename: V
authors.initials: V
authors.lastname: Palaniappan
authors.forename: D F
authors.initials: DF
authors.lastname: Bocian
mesh_terms: D001429:Bacteriochlorophylls / Q000737:chemistry / Q000235:genetics; D002338:Carotenoids / Q000737:chemistry; D004563:Electrochemistry; D045342:Light-Harvesting Protein Complexes; D015394:Molecular Structure; D010084:Oxidation-Reduction; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000235:genetics; D017354:Point Mutation; D011487:Protein Conformation; D016354:Rhodobacter capsulatus / Q000737:chemistry* / Q000235:genetics; D013059:Spectrum Analysis, Raman
chemical_list: D001429:Bacteriochlorophylls; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D002338:Carotenoids