Characterization of an FMO variant of Chlorobaculum tepidum carrying bacteriochlorophyll a esterified by geranylgeraniol.
The Fenna-Matthews-Olson light-harvesting antenna (FMO) protein has been a model system for understanding pigment-protein interactions in the energy transfer process in photosynthesis. All previous studies have utilized wild-type FMO proteins from several species. Here we report the purification and characterization of the first FMO protein variant generated via replacement of the esterifying alcohol at the C-17 propionate residue of bacteriochlorophyll (BChl) a, phytol, with geranylgeraniol, which possesses three more double bonds. The FMO protein still assembles with the modified pigment, but both the whole cell absorption and the biochemical purification indicate that the mutant cells contain a much less mature FMO protein. The gene expression was checked using qRT-PCR, and none of the genes encoding BChl a-binding proteins are strongly regulated at the transcriptional level. The smaller amount of the FMO protein in the mutant cell is probably due to the degradation of the apo-FMO protein at different stages after it does not bind the normal pigment. The absorption, fluorescence, and CD spectra of the purified FMO variant protein are similar to those of the wild-type FMO protein except the conformations of most pigments are more heterogeneous, which broadens the spectral bands. Interestingly, the lowest-energy pigment binding site seems to be unchanged and is the only peak that can be well resolved in 77 K absorption spectra. The excited-state lifetime of the variant FMO protein is unchanged from that of the wild type and shows a temperature-dependent modulation similar to that of the wild type. The variant FMO protein is less thermally stable than the wild type. The assembly of the FMO protein and also the implications of the decreased FMO/chlorosome stoichiometry are discussed in terms of the topology of these two antennas on the cytoplasmic membrane.
Department of Biology, Washington University, St. Louis, Missouri 63130, USA.
Jianzhong
J
Wen
Jiro
J
Harada
Kenny
K
Buyle
Kevin
K
Yuan
Hitoshi
H
Tamiaki
Hirozo
H
Oh-Oka
Richard A
RA
Loomis
Robert E
RE
Blankenship
D001426:Bacterial Proteins / Q000235:genetics*; D025541:Bacteriochlorophyll A / Q000378:metabolism*; D019414:Chlorobi; D004224:Diterpenes / Q000378:metabolism*; D004735:Energy Transfer; D004951:Esterification; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D014644:Genetic Variation; D045342:Light-Harvesting Protein Complexes / Q000235:genetics*; D010788:Photosynthesis; D010836:Phytol; D055550:Protein Stability
D001426:Bacterial Proteins; D025541:Bacteriochlorophyll A; D004224:Diterpenes; C086320:FMO bacteriochlorophyll protein, Bacteria; D045342:Light-Harvesting Protein Complexes; D010836:Phytol; C017338:geranylgeraniol