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1.
Chikashi Nakamura Miki Hasegawa Kazumi Shimada Makoto Shirai Jun Miyake 《Biotechnology and Bioprocess Engineering》2000,5(6):413-417
In this study, a direct detection system for triazine derivative herbicides was developed using the photosynthetic reaction
center (RC) from the purple bacterium,Rhodobacter sphaeroides, and surface plasmon resonance (SPR) apparatus. The histidine-tagged RCs were immobilized on an SPR gold chip using nickel-nitrilotriacetic
acid groups as a binder for one of the triazine herbicide, atrazine. The SPR responses were proportional to the sample concentrations
of atrazine in the range 0.1–1 μg/mL. The sensitivity of the direct detection of atrazine using the RC-assembled sensor chip
was higher than that using the antibody-immobilized chip. The other types of herbicides, DCMU or MCPP, were not detected with
such high sensitivity. The results indicated the high binding selectivity of the RC complex. 相似文献
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T Nozawa J T Trost T Fukada M Hatano J D McManus R E Blankenship 《Biochimica et biophysica acta》1987,894(3):468-476
Reaction centers were purified from the thermophilic purple sulfur photosynthetic bacterium Chromatium tepidum. The reaction center consists of four polypeptides L, M, H and C, whose apparent molecular masses were determined to be 25, 30, 34 and 44 kDa, respectively, by polyacrylamide gel electrophoresis. The heaviest peptide corresponds to tightly bound cytochrome. The tightly bound cytochrome c contains two types of heme, high-potential c-556 and low-potential c-553. The low-potential heme is able to be photooxidized at 77 K. The reaction center exhibits laser-flash-induced absorption changes and circular dichroism spectra similar to those observed in other purple photosynthetic bacteria. Whole cells contain both ubiquinone and menaquinone. Reaction centers contain only a single active quinone; chemical analysis showed this to be menaquinone. Reaction center complexes without the tightly bound cytochrome were also prepared. The near-infrared pigment absorption bands are red-shifted in reaction centers with cytochrome compared to those without cytochrome. 相似文献
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Tsunenori Nozawa Jeffrey T. Trost Taisei Fukada Masahiro Hatano James D. McManus Robert E. Blankenship 《BBA》1987,894(3):468-476
Reaction centers were purified from the thermophilic purple sulfur photosynthetic bacterium Chromatium tepidum. The reaction center consists of four polypeptides L, M, H and C, whose apparent molecular masses were determined to be 25, 30, 34 and 44 kDa, respectively, by polyacrylamide gel electrophoresis. The heaviest peptide corresponds to tightly bound cytochrome. The tightly bound cytochrome c contains two types of heme, high-potential c-556 and low-potential c-553. The low-potential heme is able to be photooxidized at 77 K. The reaction center exhibits laser-flash-induced absorption changes and circular dichroism spectra similar to those observed in other purple photosynthetic bacteria. Whole cells contain both ubiquinone and menaquinone. Reaction centers contain only a single active quinone; chemical analysis showed this to be menaquinone. Reaction center complexes without the tightly bound cytochrome were also prepared. The near-infrared pigment absorption bands are red-shifted in reaction centers with cytochrome compared to those without cytochrome. 相似文献
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Masuda S Yoshida M Nagashima KV Shimada K Matsuura K 《The Journal of biological chemistry》1999,274(16):10795-10801
The nucleotide sequence of the puf operon, which contains the genes encoding the B870 light-harvesting protein and the reaction center complex of the purple photosynthetic bacterium, Rhodovulum sulfidophilum, was determined. The operon, which consisted of six genes, pufQ, pufB, pufA, pufL, pufM, and pufC, is a new variety in photosynthetic bacteria in the sense that pufQ and pufC coexist. The amino acid sequence of the cytochrome subunit of the reaction center deduced from the pufC sequence revealed that this cytochrome contains only three possible heme-binding motifs; the heme-1-binding motif of the corresponding tetraheme cytochrome subunits was not present. This is the first exception of the "tetraheme" cytochrome family in purple bacteria and green filamentous bacteria. The pufC sequence also revealed that the sixth axial ligands to heme-1 and heme-2 irons were not present in the cytochrome either. This cytochrome was actually detected in membrane preparation as a 43-kDa protein and shown to associate functionally with the photosynthetic reaction center as the immediate electron donor to the photo-oxidized special pair of bacteriochlorophyll. This new cytochrome should be useful for studies on the role of each heme in the cytochrome subunit of the bacterial reaction center and the evolution of proteins in photosynthetic electron transfer systems. 相似文献
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The reaction centers (RCs) from several species of a purple photosynthetic bacterium, Rhodopseudomonas palustris, were first isolated by ammonium-sulfate fractionation of the isolated core complexes, and were successfully purified by anion-exchange and gel-filtration chromatography as well as sucrose-density gradient centrifugation. The RCs were characterized by spectroscopic and biochemical analyses, indicating that they were sufficiently pure and had conserved their redox activity. The pigment composition of the purified RCs was carefully analyzed by LCMS. Significant accumulation of both bacteriochlorophyll(BChl)-a and bacteriopheophytin(BPhe)-a esterified with various isoprenoid alcohols in the 17-propionate groups was shown in RCs for the first time. Moreover, a drastic decrease in BPhe-a with the most dehydrogenated and rigid geranylgeranyl(GG) ester was observed, indicating that BPhe-a in RC preferably took partially hydrogenated and flexible ester groups, i.e. dihydro-GG and tetrahydro-GG in addition to phytyl. Based on the reported X-ray crystal structures of purple bacterial RCs, the meaning of flexibility of the ester groups in BChl-a and BPhe-a as the cofactors of RCs is proposed. 相似文献
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Nobel lecture. The photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis. 总被引:23,自引:5,他引:18
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In our lectures we first describe the history and methods of membrane protein crystallization, before we show how the structure of the photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis was solved. Then the structure of this membrane protein complex is correlated with its function as a light-driven electron pump across the photosynthetic membrane. Finally we draw conclusions on the structure of the photosystem II reaction centre from plants and discuss the aspects of membrane protein structure. Sections 1 (crystallization), 4 (conclusions on the structure of photosystem II reaction centre and evolutionary aspects) and 5 (aspects of membrane protein structure) were presented and written by H.M., Sections 2 (determination of the structure) and 3 (structure and function) by J.D. We have arranged the paper in this way in order to facilitate continuous reading. 相似文献
13.
A neutral serine protease was purified as a homogeneous protein from the culture broth of photosynthetic bacterium T-20 by sequential chromatographies on columns of DEAE-cellulose, Toyopearl HW 55F, hydroxyapatite, and CM-cellulose. The molecular weight was estimated to be approximately 44,000 by SDS-PAGE, while the value of approximately 80,000 was obtained when the Hedrick-Smith method was used; this suggested that the enzyme consists of two identical subunits. The isoelectric point was determined to be 6.3 by isoelectric focusing. The enzyme had a pH optimum at 7.8. Maximal enzyme activity was detected at 50°C, and the activity was stable up to 50°C for 5 min at pH 7.0–7.2. The substrate specificity of the protease was investigated with a series of synthetic peptidyl-p-nitroanilide. The best substrate examined was Suc-Ala-Ala-Pro-Phe-pNA. The protease activity was inhibited by various inhibitors of serine protease such as chymostatin, PMSF, and DFP. EDTA, which is an inhibitor of metal protease, also inhibited the protease activity, whereas inhibitors of thiol and aspartic proteases had no significant effect. 相似文献
14.
Kimura Y Alric J Verméglio A Masuda S Hagiwara Y Matsuura K Shimada K Nagashima KV 《The Journal of biological chemistry》2007,282(9):6463-6472
A new type of membrane-bound cytochrome c was found in a marine purple photosynthetic bacterium, Rhodovulum sulfidophilum. This cytochrome c was significantly accumulated in cells growing under anaerobic photosynthetic conditions and showed an apparent molecular mass of approximately 100 kDa when purified and analyzed by SDS-PAGE. The midpoint potential of this cytochrome c was 369 mV. Flash-induced kinetic measurements showed that this new cytochrome c can work as an electron donor to the photosynthetic reaction center. The gene coding for this cytochrome c was cloned and analyzed. The deduced molecular mass was nearly equal to 50 kDa. Its C-terminal heme-containing region showed the highest sequence identity to the water-soluble cytochrome c(2), although its predicted secondary structure resembles that of cytochrome c(y). Phylogenetic analyses suggested that this new cytochrome c has evolved from cytochrome c(2). We, thus, propose its designation as cytochrome c(2m). Mutants lacking this cytochrome or cytochrome c(2) showed the same growth rate as the wild type. However, a double mutant lacking both cytochrome c(2) and c(2m) showed no growth under photosynthetic conditions. It was concluded that either the membrane-bound cytochrome c(2m) or the water-soluble cytochrome c(2) work as a physiological electron carrier in the photosynthetic electron transfer pathway of Rvu. sulfidophilum. 相似文献
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Fathir Insan Tanaka Kohki Yoza Kenji Kojima Akihiko Kobayashi Masayuki Wang Zheng-Yu Lottspeich Friedrich Nozawa Tsunenori 《Photosynthesis research》1997,51(1):71-82
The complete nucleotide sequences of the genes coding for L, M protein subunits and part of cytochrome subunit of the photosynthetic reaction center were determined for the thermophilic purple sulfur bacterium t Chromatium tepidum (t Chr. tepidum) which belongs to the subclass. The DNA fragments with 860 bp and 1900 bp were amplified by the Polymerase Chain Reaction (PCR) with the primers designed on the basis of amino acid sequences according to chemical sequence analysis of the proteins. The deduced amino acid sequences of these genes showed a significantly high degree of homology with those from purple non-sulfur bacteria. The L subunit consisted of 280 amino acids and had a molecular mass of 31,393. The M subunit consisted of 324 amino acids and had a molecular mass of 36,299. The aligned sequences of the L subunits of other purple bacterial reaction center polypeptides, showed the insertion of 8 amino acids in t Chr. tepidum in the connection of the first and second membrane-spanning helices different from those of purple non-sulfur bacteria. The aligned sequences of the L, M and cytochrome subunits were compared with other species and discussed in terms of phylogenetic trees. 相似文献
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Origin of mitochondria by intracellular enslavement of a photosynthetic purple bacterium 总被引:2,自引:0,他引:2
Cavalier-Smith T 《Proceedings. Biological sciences / The Royal Society》2006,273(1596):1943-1952
Mitochondria originated by permanent enslavement of purple non-sulphur bacteria. These endosymbionts became organelles through the origin of complex protein-import machinery and insertion into their inner membranes of protein carriers for extracting energy for the host. A chicken-and-egg problem exists: selective advantages for evolving import machinery were absent until inner membrane carriers were present, but this very machinery is now required for carrier insertion. I argue here that this problem was probably circumvented by conversion of the symbiont protein-export machinery into protein-import machinery, in three phases. I suggest that the first carrier entered the periplasmic space via pre-existing beta-barrel proteins in the bacterial outer membrane that later became Tom40, and inserted into the inner membrane probably helped by a pre-existing inner membrane protein, thereby immediately providing the protoeukaryote host with photosynthesate. This would have created a powerful selective advantage for evolving more efficient carrier import by inserting Tom70 receptors. Massive gene transfer to the nucleus inevitably occurred by mutation pressure. Finally, pressure from harmful, non-selected gene transfer to the nucleus probably caused evolution of the presequence mechanism, and photosynthesis was lost. 相似文献
17.
《BBA》1987,890(2):127-133
A photosynthetic reaction center complex has been purified from an aerobic photosynthetic bacterium, Erythrobacter species OCh 114. The reaction center was solubilized with 0.45% lauryldimethylamine N-oxide and purified by DEAE-Sephacel column chromatography. Absorption spectra of both reduced and oxidized forms of the reaction center were very similar to those of the reaction center from Rhodopseudomonas sphaeroides R-26 except for the contributions due to cytochrome and carotenoid. 1 mol reaction center contained 4 mol bacteriochlorophyll a, 2 mol bacteriopheophytin a, 4 mol cytochrome c-554, 2 mol ubiquinone-10, and carotenoid. The reaction center consisted of four different polypeptides of 26, 30, 32 and 42 kDa. The last one retained heme c. Absorbance at 450 nm oscillated with the period of two on consecutive flashes. The light-minus-dark difference spectrum had two peaks at 450 nm and 420 nm, indicating that odd flashes generated a stable ubisemiquinone anion and even flashes generated quinol. o-Phenanthroline accelerated the re-reduction of flash-oxidized reaction centers, indicating that o-phenanthroline inhibited the electron transfer between QA and QB. The cytochrome (cytochrome c-554) in the reaction center was oxidized on flash activation. The midpoint potential of the primary electron acceptor (QA) was determined by measuring the extent of oxidation of cytochrome c-554 at various ambient potentials. The mid-point potential of QA was −44 mV, irrespective of pH between 5.5 and 5.9. 相似文献
18.
Amino acid sequence of the cytochrome subunit of the photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis 总被引:6,自引:2,他引:6
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Weyer KA Lottspeich F Gruenberg H Lang F Oesterhelt D Michel H 《The EMBO journal》1987,6(8):2197-2202
The complete nucleotide sequence of the gene encoding the cytochrome subunit of the photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis, and the derived amino acid sequence are presented. The nucleotide sequence of the gene reveals the existence of a typical bacterial signal peptide of 20 amino acid residues which is not found in the mature cytochrome subunit. The gene encoding the cytochrome subunit is preceded by the gene encoding the M subunit. Both genes overlap by 1 bp. The mature cytochrome subunit consists of 336 amino acid residues; 73% of its amino acid sequence was confirmed by protein sequencing work. The mol. wt of the cytochrome subunit including the covalently bound fatty acids and the bound heme groups is 40 500. The internal sequence homology is low, despite the symmetric structure of the cytochrome subunit previously shown by X-ray crystallographic analysis of the intact photosynthetic reaction centre. Sequence homologies to other cytochromes were not found. 相似文献
19.
Orientated binding of photosynthetic reaction centers on gold using Ni-NTA self-assembled monolayers
Trammell SA Wang L Zullo JM Shashidhar R Lebedev N 《Biosensors & bioelectronics》2004,19(12):3226-1655
Coupling of photosynthetic reaction centers (RCs) with inorganic surfaces is attractive for the identification of the mechanisms of interprotein electron transfer (ET) and for possible applications in construction of photo- and chemosensors. Here we show that RCs from Rhodobacter sphaeroides can be immobilized on gold surfaces with the RC primary donor looking towards the substrate by using a genetically engineered poly-histidine tag (His7) at the C-terminal end of the M-subunit and a Ni---NTA terminated self-assembled monolayer (SAM). In the presence of an electron acceptor, ubiquinone-10, illumination of this RC electrode generates a cathodic photocurrent. The action spectrum of the photocurrent coincides with the absorption spectrum of RC and the photocurrent decreases in response to the herbicide, atrazine, confirming that the RC is the primary source of the photoresponse. Disruption of the Ni---NTA---RC bond by imidazole leads to about 80% reduction of the photocurrent indicating that most of the photoactive protein is specifically bound to the electrode through the linker. 相似文献
20.
Reaction center (RC) complexes isolated from a Zn-bacteriochlorophyll (BChl) a-containing purple bacterium, Acidiphilium rubrum, were characterized by absorption, circular dichroism, and magnetic circular dichroism (MCD) spectroscopy. The oxidized-minus-reduced difference spectra indicated that, in this RC, the Zn-BChl a is the primary electron donor. The molecular structure of the donor was examined by measuring the ratio of the MCD intensity of the Faraday B-term (B) to the dipole strength (D). In the Q(y) region, B/D for the donor was about half those of bacteriopheophytin a and the accessory Zn-BChl a, indicating that the primary electron donor is a dimer. The magnitude of bleach of the Q(x) band was half that observed in Rhodobacter sphaeroides, suggesting the cation is localized on a single Zn-Bchl a. The absorption intensity of the higher-energy Q(y) exciton band was approximately 28% of that of the lower-energy band, and the exciton splitting was approximately 570 cm(-1), smaller than that in Rb. sphaeroides. These results indicate that, in A. rubrum, the primary electron donor is a Zn-BChl a dimer but that the interaction between the two molecules is rather weak. On the basis of these results, an adaptive strategy for changes in BChl a species is discussed from an evolutionary perspective. 相似文献