EPR characterization of genetically modified reaction centers of Rhodobacter capsulatus.
Electron paramagnetic resonance (EPR) has been used to investigate the cation and triplet states of Rhodobacter capsulatus reaction centers (RCs) containing amino acid substitutions affecting the primary donor, monomeric bacteriochlorophylls (Bchls), and the photoactive bacteriopheophytin (Bphe). The broadened line width of the cation radical in HisM200----Leu and HisM200----Phe reaction centers, whose primary donor consists of a Bchl-Bphe heterodimer, indicates a highly asymmetric distribution of the unpaired electron over the heterodimer. A T0 polarized triplet state with reduced yield is observed in heterodimer-containing RCs. The zero field splitting parameters indicate that this triplet essentially resides on the Bchl half of the heterodimer. The cation and triplet states of reaction centers containing HisM200----Gln, HisL173----Gln, GluL104----Gln, or GluL104----Leu substitutions are similar to those observed in wild type. Oligonucleotide-mediated mutagenesis has been used to change the histidine residues that are positioned near the central Mg2+ ions of the reaction center monomeric bacteriochlorophylls. Reaction centers containing serine substitutions at M180 and L153 or a threonine substitution at L153 have unaltered pigment compositions and are photochemically active. The cation and triplet states of HisL153----Leu reaction centers are similar to those observed in wild type. Triplet energy transfer to carotenoid is not observed at 100 K in HisM180----Arg chromatophores. These results have important implications for the structural requirements of tetrapyrrole binding and for our understanding of the mechanisms of primary electron transfer in the reaction center.
Bylina
E J
EJ
Department of Chemistry, Massachusetts Institute of Technology, Cambridge 02139.
Kolaczkowski
S V
SV
Norris
J R
JR
Youvan
D C
DC
D001429:Bacteriochlorophylls / Q000235:genetics* / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D045342:Light-Harvesting Protein Complexes; D009154:Mutation; D010084:Oxidation-Reduction; D010674:Pheophytins / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D011758:Pyrroles / Q000378:metabolism; D012244:Rhodospirillaceae / Q000378:metabolism*; D045725:Tetrapyrroles
D001429:Bacteriochlorophylls; D045342:Light-Harvesting Protein Complexes; D010674:Pheophytins; D045322:Photosynthetic Reaction Center Complex Proteins; D011758:Pyrroles; D045725:Tetrapyrroles; C006678:bacteriopheophytin