A reaction center-light-harvesting 1 complex (RC-LH1) from a Rhodospirillum rubrum mutant with altered esterifying pigments: characterization by optical spectroscopy and cryo-electron microscopy.
Introduction of the bchP gene from Rhodobacter sphaeroides encoding geranylgeranyl reductase into Rhodospirillum rubrum alters the esterification of the bacteriochlorophylls so that phytol is used instead of geranylgeraniol. The resulting transconjugant strain of Rs. rubrum grows photosynthetically, showing that phytolated Bchla can substitute for the native pigment in both the reaction center (RC) and the light-harvesting 1 (LH1) complexes. This genetic manipulation perturbs the native carotenoid biosynthetic pathway; several biosynthetic intermediates are assembled into the core complex and are capable of energy transfer to the bacteriochlorophylls. RC-LH1 complexes containing phytolated Bchla were analyzed by low temperature absorption and fluorescence spectroscopy and circular dichroism. These show that phytolated Bchls can assemble in vivo into the photosynthetic apparatus of Rs. rubrum and that the newly introduced phytol tail provokes small perturbations to the Bchls within their binding sites in the LH1 complex. The RC-LH1 core complex was purified from membranes and reconstituted into well ordered two-dimensional crystals with a p4212 space group. A projection map calculated to 9 A shows clearly that the LH1 ring from the mutant is composed of 16 subunits that surround the reaction center and that the diameter of this complex is in close agreement with that of the wild-type LH1 complex.
Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, United Kingdom.
Pu
P
Qian
Hugh A
HA
Addlesee
Alexander V
AV
Ruban
Peiyi
P
Wang
Per A
PA
Bullough
C Neil
CN
Hunter
D001429:Bacteriochlorophylls; D002338:Carotenoids / Q000096:biosynthesis; D002462:Cell Membrane / Q000737:chemistry; D020285:Cryoelectron Microscopy; D003460:Crystallization; D004951:Esterification; D045342:Light-Harvesting Protein Complexes; D009154:Mutation; D010088:Oxidoreductases; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D010860:Pigments, Biological / Q000737:chemistry*; D012247:Rhodospirillum rubrum / Q000737:chemistry*; D013329:Structure-Activity Relationship
D001429:Bacteriochlorophylls; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D010860:Pigments, Biological; D002338:Carotenoids; D010088:Oxidoreductases; C402429:geranylgeranyl-bacteriochlorophyll reductase