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1.
Green sulfur photosynthetic bacteria optimize their antennas, chlorosomes, especially for harvesting weak light by organizing bacteriochlorophyll (BChl) assembly without any support of proteins. As it is difficult to crystallize the organelles, a high-resolution structure of the light-harvesting devices in the chlorosomes has not been clarified. We have determined the structure of BChl c assembly in the intact chlorosomes from Chlorobium limicola on the basis of 13C dipolar spin-diffusion solid-state NMR analysis of uniformly 13C-labeled chlorosomes. About 90 intermolecular C–C distances were obtained by the simultaneous assignment of distance correlations and the structure optimization preceded by the polarization-transfer matrix analysis. An atomic structure was obtained, using these distance constraints. The determined structure of the chlorosomal BChl c assembly is built with the parallel layers of piggyback-dimers. This supramolecular structure would provide insights into the mechanism of weak-light capturing. 相似文献
2.
13C-enriched bacteriochlorophyll c (R[E, E] BChl cF) was suspended in chloroform to form an aggregate showing the Qy absorption at 705 nm. (1) The aggregate exhibited several largely split 13C-NMR signals suggesting the presence of non-equivalent BChl c molecules in the form of the piggyback dimer. (2) Changes in the 13C chemical shifts were traced when methanol was titrated to dissolve the aggregate, and the aggregation shifts (in reference to the monomeric state) were determined as a function of the amount of methanol titrated, and they were analyzed empirically. (3) The ring-current effects were calculated based on the loop-current approximation, and the results were compared with the observed aggregation shifts for 13C and 1H nuclei (the 1H aggregation shifts were determined by extrapolation of the data taken from Mizoguchi, T.; Limantara, L.; Matsuura, K.; Shimada, K.; Koyama, Y. J Mol Structure 1996, 379, 249-265). The results showed that the assembly of two straight columns consisting of the piggyback dimer stacked in the antiparallel orientation is the best choice as a model for the B705 aggregate. (4) Three-dimensional F1 13C-edited F3 13C-filtered heteronuclear single-quantum nuclear-Overhauser-effect spectroscopy was applied to the aggregate consisting of a 1:1 mixture of 13C-labeled and unlabeled BChl c in order to selectively detect the intermolecular 1H-1H NOE correlations. The NOE correlations were explained in terms of a straight column, supporting the above model. 相似文献
3.
Pigments including bacteriochlorophyll (BChl) c, carotenoids, and a trace of BChl a together with a lipid, monogalactosyl diglyceride (MGDG), were extracted with chloroform/methanol (1:1 v/v) from an aqueous suspension (50 mM Tris-HCl, pH 8.0) of chlorosomes from Chlorobium limicola; other lipids and proteins were left behind in the aqueous layer by funnel separation. The chloroform layer was dried by purging N2 gas, dissolved in methanol, and rapidly injected into the aqueous layer to reassemble chlorosomes. This technique has been developed to replace one-half of the inherent 12C-BChl c by 13C-BChl c to identify the intermolecular 13C...13C magnetic dipole correlation peaks (that are supposed to reduce their intensities to one-fourth by reducing the 13C-BChl c concentration into one-half) and to determine the structure of BChl c aggregates in the rod elements by means of solid-state NMR spectroscopy. The isotopically replaced chlorosomes were characterized (1) by sucrose density gradient centrifugation, zeta potential measurement, electron microscopy, and dynamic light scattering measurement to determine the morphology of chlorosomes, (2) by 13C NMR spectroscopy, electronic absorption and circular dichroism spectroscopies, and low-angle X-ray diffraction to determine the pigment assembly in the rod elements, and (3) by subpicosecond time-resolved absorption spectroscopy to determine the excited-state dynamics in the pigment assembly. The results characterized the reassembled chlorosomes to have (1) similar but longer morphological structures, (2) almost the same pigment assembly in the rod elements, and (3) basically the same excited-state dynamics in the pigment assembly. 相似文献
4.
High activities of ATP sulfurylase were found in the soluble protein fraction of two Chlorobium limicola strains, whereas ADP sulfurylase was absent. ATP sulfurylase was partially purified and characterized. It was a stable soluble enzyme with a molecular weight of 230,000, buffer-dependent pH optima at 8.6 and 7.2 and an isoelectric point at pH 4.8. No physiological inhibitor was found. Inhibition was observed with p-CMB and heavy metals. Sulfur compounds had no effect on enzyme activity. The stoichiometry of the reaction was proven. In contrast, an ADP sulfurylase, but no ATP sulfurylase, was found in Chlorobium vibrioforme. This enzyme was very labile with a molecular weight of about 120,000 and buffer-dependent pH optima at 9.0 and 8.5. Under test conditions the apparent K
m value was determined to be 0.28 mM for adenylyl sulfate and 8.0 mM for phosphate.Abbreviations APS
adenylyl sulfate
- p-CMB
parachloromercuribenzoate
- PPi
inorganic pyrophosphate 相似文献
5.
In this paper, a three-dimensional (3D) NMR-based approach for the determination of the fold of moderately sized proteins by solid-state magic-angle spinning (MAS) NMR is presented and applied to the alpha-spectrin SH3 domain. This methodology includes the measurement of multiple (13)C-(13)C distance restraints on biosynthetically site-directed (13)C-enriched samples, obtained by growing bacteria on [2-(13)C]glycerol and [1,3-(13)C]glycerol. 3D (15)N-(13)C-(13)C dipolar correlation experiments were applied to resolve overlap of signals, in particular in the region where backbone carbon-carbon correlations of the C(alpha)-C(alpha), CO-CO, C(alpha)-CO, and CO-C(alpha) type appear. Additional restraints for confining the structure were obtained from phi and psi backbone torsion angles of 29 residues derived from C(alpha), C(beta), CO, NH, and H(alpha) chemical shifts. Using both distance and angular restraints, a refined structure was calculated with a backbone root-mean-square deviation of 0.7 A with respect to the average structure. 相似文献
6.
Chlorobium limicola forma thiosulfatophilum: Biocatalyst in the Production of Sulfur and Organic Carbon from a Gas Stream Containing H2S and CO2 总被引:3,自引:2,他引:1
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Chlorobium limicola forma thiosulfatophilum (ATCC 17092) was grown in a 1-liter continuously stirred tank reactor (800-ml liquid volume) at pH 6.8, 30°C, saturated light intensity, and a gas flow rate of 23.6 ml/min from a gas cylinder blend consisting of 3.9 mol% H2S, 9.2 mol% CO2, 86.4 mol% N2, and 0.5 mol% H2. This is the first demonstration of photoautotrophic growth of a Chlorobium sp. on a continuous inorganic gas feed. A significant potential exists for applying this photoautotrophic process to desulfurization and CO2 fixation of gases containing acidic components (H2S and CO2). 相似文献
7.
A. García-Granados A. Martínez A. Molina M.E. Onorato M. Rico A.Saenz de Buruaga J.M.Saenz de Buruaga 《Phytochemistry》1985,24(8):1789-1792
Several ent-13-epi-manoyl oxides and a new natural product, ent-8α-hydroxylabda-13(16),14-dien-19-oic acid, have been isolated from Sideritis varoi subsp. cuatrecasasii. In addition, a study has been made of some natural and semisynthetic ent-13-epi-manoyl oxides functionalized at C-12. 相似文献
8.
Oh-oka Hirozo; Kakutani Saki; Matsubara Hiroshi; Malkin Richard; Itoh Shigeru 《Plant & cell physiology》1993,34(1):93-101
The photoactive reaction center (RC) complex from the greensulfur bacterium Chlorobium limicola f. thiosulfatophilum, strainLarsen, was isolated after solubilization and ammonium sulfatefractionation followed by ion-exchange chromatography. The spectrumof the complex was almost identical with that of the similarRC complex isolated by Feiler et al. [(1992) Biochemistry 31:26082614] except for the presence of cytochrome c551instead of c553 in the latter study. A molecular ratio of BChla to P840 of the isolated RC complex was assayed to be 2535.SDSPAGE analysis revealed that the isolated complex containedthree major polypeptides with apparent molecular masses of 68,41 and 21 kDa, respectively. The 21-kDa polypeptide was identifiedto be a heme-binding protein by staining the gel for peroxidaseactivity. The cytochrome c551 was oxidized by flash light ina biphasic manner with half times of 90 and 390 µs, respectively,that coincided with the reduction half times of P840+. Threedistinct iron-sulfur centers assigned to FA, FB and Fx, respectively,from their g-values were detected by EPR spectroscopy at cryogenictemperature. These results suggest that the present preparationcontains a minimal functional unit of the RC of this bacterium,and that this complex appears to lie on a evolutionary linebetween RC's of purple bacteria and photosystem I. (Received August 18, 1992; Accepted October 28, 1992) 相似文献
9.
Study of phospholipid structure by 1H, 13C, and 31P dipolar couplings from two-dimensional NMR.
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Various motionally averaged 31P-1H, 13C-1H, 1H-1H, and 31P-13C dipolar couplings were measured for natural-abundance and unoriented phosphocholine in the L alpha phase. The couplings were obtained and assigned by a variety of advanced and partly novel two-dimensional solid-state NMR experiments. Whereas 31P-1H and 31P-13C dipolar couplings provide long-range structural constraints, geminal 1H-1H couplings and the signs of 13C-1H couplings are important new elements in a segmental order-tensor analysis of the lipid headgroup and glycerol backbone. The implications of these measured dipolar couplings for the conformational exchange of the lipid headgroup and the bending of the headgroup from the glycerol backbone are discussed. These dipolar couplings are also analyzed semiquantitatively in terms of the segmental order tensor. 相似文献
10.
A M van den Besselaar B de Kruijff H van den Bosch L L van Deenen 《Biochimica et biophysica acta》1979,555(2):193-199
1. The transverse distribution of 1-palmitoyl-sn-glycero-3-phospho-N-[Me-13C]-choline in vitro incorporated in sarcoplasmic reticulum has been measured by means of 13C NMR and DyCl3 as an impermeable shift reagent. 2. Lysophosphatidylcholine added to the membranes equilibrates within 30 min at 20 degrees C between outer and inner membrane leaflet so that 42% is located in the inner leaflet. 3. Lysophosphatidylcholine diffuses back from the inner leaflet to the outer upon lysophospholipase action on the outer lysophosphatidylcholine pool. 相似文献
11.
Chlorobium limicola forma thiosulfatophilum (ATCC 17092) was grown in a 1-liter continuously stirred tank reactor (800-ml liquid volume) at pH 6.8, 30 degrees C, saturated light intensity, and a gas flow rate of 23.6 ml/min from a gas cylinder blend consisting of 3.9 mol% H(2)S, 9.2 mol% CO(2), 86.4 mol% N(2), and 0.5 mol% H(2). This is the first demonstration of photoautotrophic growth of a Chlorobium sp. on a continuous inorganic gas feed. A significant potential exists for applying this photoautotrophic process to desulfurization and CO(2) fixation of gases containing acidic components (H(2)S and CO(2)). 相似文献
12.
Characterization of Chlorobium tepidum chlorosomes: a calculation of bacteriochlorophyll c per chlorosome and oligomer modeling
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Montaño GA Bowen BP LaBelle JT Woodbury NW Pizziconi VB Blankenship RE 《Biophysical journal》2003,85(4):2560-2565
The bacteriochlorophyll (Bchl) c content and organization was determined for Chlorobium (Cb.) tepidum chlorosomes, the light-harvesting complexes from green photosynthetic bacteria, using fluorescence correlation spectroscopy and atomic force microscopy. Single-chlorosome fluorescence data was analyzed in terms of the correlation of the fluorescence intensity with time. Using this technique, known as fluorescence correlation spectroscopy, chlorosomes were shown to have a hydrodynamic radius (Rh) of 25 +/- 3.2 nm. This technique was also used to determine the concentration of chlorosomes in a sample, and pigment extraction and quantitation was used to determine the molar concentration of Bchl c present. From these data, a number of approximately 215,000 +/- 80,000 Bchl c per chlorosome was determined. Homogeneity of the sample was further characterized by dynamic light scattering, giving a single population of particles with a hydrodynamic radius of 26.8 +/- 3.7 nm in the sample. Tapping-mode atomic force microscopy (TMAFM) was used to determine the x,y,z dimensions of chlorosomes present in the sample. The results of the TMAFM studies indicated that the average chlorosome dimensions for Cb. tepidum was 174 +/- 8.3 x 91.4 +/- 7.7 x 10.9 +/- 2.71 nm and an overall average volume 90,800 nm(3) for the chlorosomes was determined. The data collected from these experiments as well as a model for Bchl c aggregate dimensions was used to determine possible arrangements of Bchl c oligomers in the chlorosomes. The results obtained in this study have significant implications on chlorosome structure and architecture, and will allow a more thorough investigation of the energetics of photosynthetic light harvesting in green bacteria. 相似文献
13.
Kinetic parameters of human carbonic anhydrase B as determined from NMR linewidths of 13 C in CO 2 and HCO 3 - 总被引:1,自引:0,他引:1
The linewidths of the 13C NMR signals of CO2 and , in equilibrium aqueous solutions containing small amounts of carbonic anhydrase, are determined mainly by the rate of enzyme-induced interconversion of CO2 and . We have measured these linewidths in unbuffered solutions of human carbonic anhydrase B for several values of [CO2], at 25°C as a function of pH. From a least-squares analysis of the data, using the equations relating the linewidths to the enzyme kinetics, we have obtained values for the kinetic (Michaelis-Menten) parameters that characterize this interconversion. These preliminary results are in approximate agreement with published values for highly buffered solutions. Additionally, the results confirm that the product of the hydration reaction, and the substrate for the dehydration, is the neutral molecule H2CO3. 相似文献
14.
15.
We report on the conformation of a tetrasaccharide fragment in the repeating subunit of the cell wall polysaccharide of Streptococcus mitis J22, a receptor for the lectin of Actinomyces viscosus T14V in a bacterial coaggregation that is important in the ecological interactions of oral bacteria. Although there is considerable overlap of the 1H-nmr signals, some cross peaks can be extracted from conventional two-dimensional nuclear Overhauser effect spectroscopy (NOESY) data on the polysaccharide. These data cannot be fit to a single conformation of the tetrasaccharide fragment. Therefore we have prepared a polysaccharide sample fully enriched in 13C from which we have determined accurate NOESY cross-peak volumes in a three-dimensional heteronuclear-resolved spectrum that allows accurate determination of many more NOESY cross peaks than does conventional two-dimensional spectroscopy. We have also used the 13C enriched polysaccharide to measure accurate values of long-range 13C-1H coupling constants that can be correlated with glycosidic dihedral angles. Molecular modeling calculations on the polysaccharide fragment, including molecular dynamics simulations, identify multiple low-energy conformations. This result is to be contrasted with previous calculations on blood group oligosaccharides in our laboratory using similar methods that showed relatively rigid conformations with little flexibility of the glycosidic linkages. The present NOESY and 3JCH data can be reconciled with a model for the antigenic tetrasaccharide in which three distinct conformations are in fast exchange. We propose that some carbohydrate epitopes such as those of the blood group oligosaccharides are relatively rigid while others such as the tetrasaccharide fragment in these studies exhibit much greater flexibility. © 1996 John Wiley & Sons, Inc. 相似文献
16.
Rotational diffusion and intermolecular collisions of a spin labeled alpha-helical peptide determined by electron spin echo spectroscopy.
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Short peptides that are composed mainly of alanine have recently been shown to form alpha-helices in aqueous solution at low temperature (Marqusee, S., and R. L. Baldwin. 1987. Proc. Natl. Acad. Sci. 84:8898-8902; Marqusee, S., V. H. Robbins, and R. L. Baldwin. 1989. Proc. Natl. Acad. Sci. USA. 86:5286-5290). These peptides are excellent models for probing structure and dynamics in isolated helical domains. In previous work we have designed and synthesized spin labeled analogs of these helix-forming peptides and we have shown that these analogs retain the folding characteristics of the parent peptide (Todd, A. P., and G. L. Millhauser. 1991. Biochemistry. 30:5515-5523). Using conventional continuous wave electron spin resonance (CW ESR) we have further shown that local motion is more pronounced near the helix amino terminus than in the central region as the peptide is thermally unfolded (Miick, S. M., A. P. Todd, and G. L. Millhauser. 1991. Biochemistry. 30:9498-9503). In this present work we use electron spin echo (ESE) spectroscopy to further refine our understanding of the solution dynamics of the 3K-8 peptide, which is a 16-mer with a nitroxide spin label attached at position 8. We find that the spin echo decays are well described by a single exponential function and that the determined correlation times are close to those previously derived from CW experiments. Variable concentration ESE experiments have directly revealed Heisenberg spin exchange (HSE) interactions and we find that the interpeptide collision rate is near to that expected for a free species in solution.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
17.
Base treatment of O-benzyl protected C-2- or C-4-ulopyranosyl compounds (4 alpha, 4 beta, and 11) by either 10% Et(3)N or 1% K(2)CO(3) in MeOH initiated a beta elimination to afford alpha,beta-unsaturated C-ulopyranosyl compounds (5 alpha, 5 beta, and 12), which further rearranged in a stereocontrolled manner to multifuctionalized alpha,beta-cyclopentenones (6 and 14) in 70-80% yield. Both C-alpha- and C-beta-2-ulosides (5 alpha and 5 beta) produced the same cyclopentenone 6, indicating that a 1,2-enolate is formed prior to the cleavage of the C-5--O bond. Because 6 is racemic, it was probably formed by the intramolecular cycloaldolization of two equally populated enantiomeric intermediates. When treated with 90% Et(3)N in MeOH, 5 alpha yielded almost exclusively 15 (isomer of 6), which was formed by a migration of the double bond in 5 alpha during the previously described rearrangement. Thus either 6 or 15 was the major product, depending on the base used. 相似文献
18.
The reaction of N-[1-13C] acetylimidazole with cytochrome c and guanidinated cytochrome c was evaluated as a means of introducing NMR-detectable groups as conformation-dependent probes. Resonances from both N-[1-13C]acetyl lysyl and O-[1-13C]acetyl tyrosyl groups were observed when ferricytochrome c was acetylated. However, only O-[1-13C]acetyl tyrosyl resonances were seen with acetylated guanidinated ferricytochrome c. Chemical shifts of the four O-[1-13C]acetyl tyrosyl groups were conformation dependent and ranged from 172 to 176 ppm. A convenient method for the preparation of N-[1-13C]acetylimidazole is described. 相似文献
19.
Soil aggregates in a tropical deciduous forest: effects on C and N dynamics, and microbial communities as determined by t-RFLPs 总被引:1,自引:0,他引:1
Ana M. Noguez Ana E. Escalante Larry J. Forney Maribel Nava-Mendoza Irma Rosas Valeria Souza Felipe García-Oliva 《Biogeochemistry》2008,89(2):209-220
The aim of this study was to analyze C and N dynamics, as well as, soil bacterial community structure within soil micro- and
macro-aggregates in a tropical deciduous forest in México. We measured, for three landscape positions and three seasons of
the year: total, microbial and available forms of C and N; potential C and N mineralization; and soil bacterial communities
by using t-RFLPs. The highest total C concentrations were found in the north-slopes and in the dry season (DS) samples. In
general, micro-aggregates had higher concentrations than macro-aggregates of available C and N forms, and microbial C. Similarly,
micro-aggregates had the highest potential C mineralization and net N mineralization. We detected 149 different OTUs (operational
taxonomic units) from which 50% was shared by the two aggregate size fractions, 25% was exclusive to micro-aggregates and
the 25% left was found only in macro-aggregates. Top-hills were richer in OTUs than north and south-slopes. The Unweighted
Pair Group Method with Arithmetic mean (UPGMA) analysis indicated clear differences in community composition between the two
aggregate size-fractions in relation to the presence of OTUs. These results suggest that the main difference between micro-
and macro-aggregates is due to the community structure within each soil fraction and this difference could affect soil nutrients
dynamics. 相似文献
20.
Ling Han Magnus Doverskog Sven-Olof Enfors Lena H?ggstr?m 《Journal of biotechnology》2002,92(3):237-249
Addition of selected amino acids could be a means to improve production of recombinant proteins in industrial processes. We found that glycine increased the maximum specific growth rate of Escherichia coli from 0.67 to 0.78 h(-1), and the cell yield from 0.57 to 0.98 g dry weight per g substrate, when supplemented to batch cultures in a glucose-mineral medium. Maximum effect occurred at pH 6.8, at a glycine concentration of 6-12 mmol l(-1), and at cell densities below 1.15 g dry weight l(-1) (0D(610).3). When glycine was added to a culture at a cell density of 1.15 g l(-1) or above, no growth promoting effect of glycine was seen. The 'glycine effect' was not due to CO(2) produced by the glycine cleavage system (GCV), and the lack of effect at higher cell densities was not masked by acetate accumulation, but coincided with increased acetate production. The metabolism of glycine was further investigated in cultures supplied with [2-(13)C] labelled glycine, and the redistribution of label in the [1-(13)C], [2-(13)C], and [1,2-(13)C] isotopomeres of excreted acetate was analysed by 13C NMR. The NMR data revealed that very little degradation of glycine occurred at cell densities below 1.15 g l(-1). Simultaneously the biosynthesis of serine and glycine was repressed as judged by the absence of [2-(13)C] acetate, implying that added glycine was used as a source of glycine, serine, one-carbon units, and threonine. At cell densities above 1.15 g l(-1), 53% of the consumed glycine carbon was excreted as acetate. Degradation of glycine was associated with an increased uptake rate, cleavage by GCV, and degradation of both glycine-derived serine, and glucose-derived serine to pyruvate. This switch in metabolism appears to be regulated by quorum sensing. 相似文献