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Biomedical subjects

G Drews

Publications and source records attributed to G Drews.

At least 109 records · Page 6Linked to original sources
PubMed1983

Primary structure of the peptidoglycan from the unicellular cyanobacterium Synechocystis sp. strain PCC 6714.

A peptidoglycan fraction free of non-peptidoglycan components was isolated from the unicellular cyanobacterium Synechocystis sp. strain PCC 6714. Hydrofluoric acid treatment (48%, 0 degrees C, 48 h) cleaved off from the peptidoglycan non-peptidoglycan glucosamine, mannosamine, and mannose. The purified peptidoglycan consists of N-acetyl muramic acid, N-acetyl glucosamine, L-alanine, D-alanine, D-glutamic acid, and meso-diaminopimelic acid in approximately equimolar amounts. At least partial amidation of carboxy groups in the peptide subunits is indicated. Peptide analyses and 2,4-dinitrophenyl studies of partial acid hydrolysates revealed the structure of the Synechocystis sp. strain PCC 6714 peptidoglycan to belong to the A1 gamma type (direct cross-linkage) of peptidoglycan classification. The degree of cross-linkage is about 56% and thus is in the range of that found in gram-positive bacteria. Some of the peptide units are present as tripeptides lacking the carboxy-terminal D-alanine.

Amino Acid Sequence↗
PubMed1983

Chemical cross-linking studies of the light-harvesting pigment-protein complex B800-850 of Rhodopseudomonas capsulata.

The spatial relationship of the three polypeptides contained in the B800-850 light-harvesting complex of Rhodopseudomonas capsulata has been studied with chemical cross-linking of crude membrane preparations of the phototrophic negative mutant strain Y5. Samples were cross-linked with the cleavable reagent dithiobis (succinimidyl propionate) (1.1 nm chain length) and analyzed by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. Membranes labelled with 14C-amino acids were used to determine the compositional stoichiometry of cross-linked products. It was found that the two polypeptides with an apparent Mr of 8000 and 10 000, respectively, that are associated with the pigments bacteriochlorophyll a and carotenoid form homooligomers as well as heterooligomers. The data support the idea that these polypeptides are closely arranged in clusters probably containing at least four of each species. The third subunit with an Mr of 14 000, which is not associated with pigments, was found to be most susceptible to cross-linking and formed homooligomers but no heterooligomers with the other two subunits, and is thus likely to be loosely attached to these clusters. Comparative studies with the phototrophic positive wild type strain indicated that the results found with the phototrophic negative mutant strain Y5 reflect the organization of the B800-850 complex in the membrane of Rhodopseudomonas capsulata. Studies with the isolated B800-850 complex revealed that the sterical arrangement of the three constituent polypeptides in dodecyl dimethylamine-N-oxide containing solutions must be very similar to that in the membrane.

Bacterial Proteins↗
PubMed1982

The polypeptide components from light-harvesting pigment-protein complex II (B800-850) of Rhodopseudomonas capsulata. Solubilization, purification and sequence studies.

A new procedure for isolation, purification and identification of the three polypeptides of the membrane-bound light-harvesting complex II (B800-850) of Rhodopseudomonas capsulata has been developed. The polypeptides were extracted from crude intracytoplasmic membranes with chloroform/methanol/ammonium acetate and separated by chromatography on Sephadex LH60. The peak fractions were transferred to solvents of different polarity and separated by gel filtration or ion-exchange chromatography. The three major polypeptides isolated by this two-step chromatography were found to be homogenous and identical with the three polypeptides of the light-harvesting complex II, as judged by amino acid analysis and N-terminal sequence determination. Contaminating minor polypeptides, of which the functions are unknown, were different from the polypeptides of the B800-850 complex studied by the same criteria.

Amino Acid Sequence↗
PubMed1981

Linear and circular dichroism of membranes from Rhodopseudomonas capsulata.

Absorption, linear dichroism and circular dichroism spectra of Rhodopseudomonas capsulata (wild-type-St. Louis strain, mutant Y5 and mutant Ala+) are particularly sensitive to the nature of the light-harvesting bacteriochlorophyll-carotenoid-protein complexes. Evidence for exciton-type interactions is seen near 855 nm in the membranes from the wild-type and from mutant Y5, as well as in an isolated B-800 + 850 light-harvesting complex from mutant Y5. The strong circular dichroism that reflects these interactions is attenuated more than 10-fold in membranes from the Ala+ mutant, which lacks both B-800 + 850 and colored carotenoids and contains only the B-875 light-harvesting complex. These results lead to the conclusion that these two light-harvesting complexes have significantly different chromophore arrangements or local environments.

Cell Membrane↗
PubMed1981

Comparative studies of two membrane fractions isolated from chemotrophically and phototrophically grown cells of Rhodopseudomonas capsulata.

Light and heavy membrane fractions have been isolated by equilibrium sucrose density centrifugation from Rhodopseudomonas capsulata 938 GCM grown aerobically in the dark (chemotrophically) and anaerobically in the light (phototrophically). The densities of the light and heavy fractions from phototrophic cells were 1.1004 to 1.1006 and 1.1478, respectively, and the densities of the light and heavy fractions from chemotrophic cells were 1.0957 to 1.0958 and 1.1315, respectively. Both fractions were active in photochemical and respiratory functions and in electron transport-coupled phosphorylation. The light membrane fraction isolated from chemotrophic cells contained the reaction center and the light-harvesting pigment-protein complex B 870, but not the variable light-harvesting complex B 800-850. A small amount of the complex B 800-850 was present in the light fraction isolated from phototrophically grown cells, but it was not energetically coupled to the photosynthetic apparatus. From inhibitor studies, difference spectroscopy, and measurement of enzyme activities it was tentatively concluded that the light membrane fraction contains only the reduced nicotinamide adenine dinucleotide-oxidizing electron transport chain having a KCN-insensitive, low-potential cytochrome c oxidase, whereas the heavy fraction contains additionally the succinate dehydrogenase and a high-potential cytochrome b terminal oxidase sensitive to KCN. The light membrane fraction was more labile than the heavy fraction in terms of phosphorylating activity.

Cell Fractionation↗
PubMed1980

Fluorescence emission by wild-type- and mutant-strains of Rhodopseudomonas capsulata.

Absorption and fluorescence emission spectra of Rhodopseudomonas capsulata, strains 37b4 (wild type), A1a+ (blue-green mutant strain), Y5 (phototroph negative, having only B-800--850 bacteriochlorophyll-carotenoid-protein complex) at 4 K, 77 K and 300 K were measured. The fluorescence emission at 890 nm of the B-870 bacteriochlorophyll band dominates the emission of other spectral forms of the strains 37b4 and A1a+, while in strain Y5 a fluorescence emission band at 865 nm of the B-850 bacteriochlorophyll dominates. Very little fluorescence was observed at 805 nm. A linear relation between relative fluorescence intensity and the exciting light intensity was observed. The integrated fluorescence yield increased as the temperature was lowered from 300 K to 4 K. The results are discussed in the light of the arrangement of pigment molecules in the membrane and the process of energy migration within the photosynthetic apparatus.

Bacteriochlorophylls↗
PubMed1980

Supramolecular organization of chlorosomes (chlorobium vesicles) and of their membrane attachment sites in Chlorobium limicola.

The photosynthetic green bacterium Chlorobium limicola 6230 has been examined by freeze-fracture electron microscopy to investigate the size, form, distribution and supramolecular architecture of its chlorosomes (chlorobium vesicles) as well as the chlorosome attachment sites on the cytoplasmic membrane. The oblong chlorosomes that underlie the cytoplasmic membrane show a considerable variation in size from about 40 X 70 nm to 100 X 260 nm and exhibit no particular orientation. The chlorosome core, which appears to be hydrophobic in nature, contains between 10 and 30 rod-shaped elements (approx. 10 nm in diameter) surrounded by an unetchable matrix. The rod elements are closely packed and extend the full length of the chlorosome. Separating the chlorosome core from the cytoplasm is a approx. 3 nm thick lipid-like envelope layer, which exhibits no substructure. A 5-6 nm thick, crystalline baseplate connects the chlorosome to the cytoplasmic membrane. The ridges of the baseplate lattice make an angle of between 40 degrees and 60 degrees with the longitudinal axis of the chlorosome and have a repeating distance of approx. 6 nm. In addition, each ridge exhibits a granular substructure with a periodicity of approx. 3.3 nm. The cytoplasmic membrane regions adjacent to the baseplates are enriched in large (greater than 9 nm) intramembrane particles, most of which belong to approx. 10 nm and approx. 12.5 nm particle size categories. Each chlorosome attachment site contains between 20 and 30 very large (greater than 12.0 nm diameter) intramembrane particles. The following interpretive model of a chlorosome is discussed in terms of biophysical, biochemical and structural information reported by others: it is proposed that the bacteriochlorophyll c (BChl c; chlorobium chlorophyll) is located in the rod elements of the core and that it is complexed with specific proteins. The cytoplasm-associated envelope layer is depicted as consisting of a monolayer of galactosyl diacylglycerol molecules. BChl alpha-protein complexes in a planar lattice configuration most likely make up the crystalline baseplate. The greater than 12-nm particles in the chlorosome attachment sites of the cytoplasmic membrane, finally, may correspond to complexes containing a reaction center and non-crystalline light-harvesting BChl alpha. The crystalline nature of the baseplate is consistent with the notion that it serves two functions: besides transferring excitation energy to the reaction centers it could also function as a distributor of this energy amongst the reaction centers.

Bacteria↗
PubMed1980

Isolation and characterization of the polypeptide components from light-harvesting pigment-protein complex B800--850 of Rhodopseudomonas capsulata.

The light-harvesting bacteriochlorophyll-carotenoid-protein complex B800--850 has been isolated from membranes of the phototroph-negative mutant strain Y5 of Rhodopseudomonas capsulata. The three polypeptides of the complex have been found to be soluble in chloroform-methanol (1:1, v/v) in the presence of 0.1 M ammonium acetate. They were extracted from the complex and separated by gel filtration on Sephadex LH-60 in the same solvent mixture. Minimum molecular weights based on amino acid composition are 12 000, 9300, and 5100. Values previously determined by sodium dodecyl sulfate/polyacrylamide gel electrophoresis are 14 000, 10 000 and 8000. The two smaller polypeptides (polarities 31% and 39%) are completely soluble in chloroform/methanol/ammonium acetate while the largest and most polar (41%) polypeptide is only partially soluble. The largest polypeptide contains no tryptophan. The middle polypeptide contains no cysteine and arginine, while the small polypeptide lacks cysteine. Methionine is shown to be the amino terminus for the small and middle polypeptides by two independent methods (Edman degradation and dansylation). Both methods also indicated that the N terminus of the 14 000 polypeptide seems to be blocked. Partial N-terminal amino acid sequences were obtained for the two smaller polypeptides. No homology between the two proteins was observed.

Amino Acids↗
PubMed1980

Cellular organization of the halophilic, phototrophic bacterium strain WS 68.

The obligatory halophilic, phototrophic bacterium, strain WS 68, grows under anaerobic conditions in the light in a mineral medium with acetate as carbon source and 6 to 20% sodium chloride, or in the same medium semiaerobically in the dark. The cells are vibrio- or spiral-shaped and show a bipolar polytrichous flagellation. The absorption spectrum of membranes, isolated from phototrophically grown cells, shows absorption maxima at 375, 490, 520, 555, 590, 800, 840-845 and 870-875 nm indicating the presence of bacteriochlorophyll a and carotenoids as pigments of the photosynthetic apparatus. The cell surface is covered with a layer of regular arranged particles (center to center distance approximately 10.5 nm). A murein (peptidoglycan) sacculus has not been detected in ultrathin sections, freeze fracture preparations and cells extracted with hot dodecyl sulfate. The intracytoplasmic membranes lie parallel to the cell envelope. The polypeptide patterns of isolated membranes and of the envelope fraction have been demonstrated. The results are discussed in the light of taxonomy and cell organization.

Bacteria↗
PubMed1980

The differentiation of the photosynthetic apparatus and the intracytoplasmic membrane in cells of Rhodopseudomonas capsulata upon variation of light intensity.

In cells of Rhodopseudomonas capsulata, grown at growth limiting light intensity (7 W x m-2), the cellular bacteriochlorophyll (Bchl) content increased 13-fold, the Bchl concentration of membranes 3.4 fold, the concentration of reaction centers in membranes 1.6-fold, the size of the photosynthetic unit twofold, the concentration of carotenoids in membranes 2.4 fold and the number of intracytoplasmic membrane vesicles, bearing the photosynthetic apparatus, 6..3 fold in comparison with cells grown at 2000 W x m-2 light intensity. Thus, the variation of incident caused mainly a variation of the amount of intracytoplasmic membrane vesicles per cell, but also a variation of the size and concentration of photosynthetic units in the membrane system.

Bacteriochlorophylls↗
PubMed1979

Effects of light intensity on membrane differentiation in Rhodopseudomonas capsulata.

Cells of Rhodopseudomonas capsulata, strain 37b4, leu-, precultivated anaerobically under low light intensity, were exposed to high light intensity (2000 W.m-2). The cells grew with a mass doubling time of 3 h. The synthesis of bacteriochlorophyll (BChl) began after two doublings of cell mass. Reaction center and light-harvesting BChl I (B-875) were the main constituents of the photosynthetic apparatus incorporated into the membrane. The size of the photosynthetic unit (total BChl/reaction center) decreased and light-harvesting BChl I became the dominating BChl species. Concomitant with the appearance of the different spectral forms of BChl the respective proteins were incorporated into the membrane, i.e. the three reaction center polypeptides, the polypeptide associated with light-harvesting BChl I, the two polypeptides associated with BChl II. A polypeptide of an apparent molecular weight of 45 000 was also incorporated. A lowering of the light intensity to 7 W.m-2 resulted in a lag phase of growth for 6 h. Afterwards, the time for doubling of cell mass was 11 h. The concentration of all three BChl complexes (reaction center, light-harvesting BChl I and II complexes)/cell and per membrane protein increased immediately. Also the size of the photosynthetic unit and the amount of intracytoplasmic membranes/cell increased. The activities of photophosphorylation, succinate dehydrogenase, NADH dehydrogenase and NADH oxidation (respiratory chain)/membrane protein are higher in membrane preparations isolated from cells grown at high light intensities than in such preparations from cells grown at low light intensities.

Bacteriochlorophylls↗
PubMed1979

The size and number of intramembrane particles in cells of the photosynthetic bacterium Rhodopseudomonas capsulata studied by freeze-fracture electron microscopy.

By freeze-fracture electron microscopy, particles have been observed on the protoplasmic leaflet (PF face) of cytoplasmic and intracytoplasmic membranes of the photosynthetic bacterium Rhodopseudomonas capsulata. The particles are present under all culture conditions of chemotrophically and phototrophically grown cells. However, the number of particles per microM2 increased significantly when the formation of the photosynthetic apparatus in the membrane is induced. Intracytoplasmic membranes, where the bulk of photosynthetic activity is localized, always have a higher density of particles than cytoplasmic membranes. Under all conditions particles with a diameter of 9.5 nm dominate. The frequency of particles with diameters greater or smaller than 9.5 nm changed with culture conditions. A comparison of biochemical and electron microscopic data have lead us to the conclusion that the particles, formed under conditions which allow the synthesis of the photosynthetic apparatus, are composed of photochemical reaction centers and antenna light-harvesting bacteriochlorophyll I (B 875)-protein complexes. The total molecular weight of these particles is calculated to be 500,000.

Anaerobiosis↗
PubMed1978

Isolation and characterization of light harvesting bacteriochlorophyll.protein complexes from Rhodopseudomonas capsulata.

The isolation of two native light harvesting bacteriochlorophyl.protein complexes from Rhodopseudomonas capsulata is described. The light harvesting bacteriochlorophyll I (B 875) has been isolated from the blue-green mutant A1a+ lacking both carotenoids and light harvesting bacteriochlorophyll II. Light harvesting bacteriochlorophyll I is associated with a protein (light harvesting band 2) of 12 000 molecular weight. Light harvesting bacteriochlorophyll II complex has been isolated from the mutant Y5 lacking a reaction center and light harvesting bacteriochlorophyll I. Light harvesting bacteriochlorphyll II (B 800 + 850) together with carotenoids is associated with two polypeptides (light harvesting bands 3 and 4) having molecular weights of about 8000 and 10 000 (sodium dodecyl sulfate polyacrylamide gel electrophoresis). A third protein (light harvesting band 1) is in the purified light harvesting II fraction (mol. wt. approx. 14 000), but not associated with bacteriochlorophyll or carotenoids. The amino acid composition of the 3 antenna pigment II proteins is given. The polarity of these proteins was found to be 48%. From the amino acid composition the following molecular weights were calculated band 1: 17 350, band 3: 13 350 and band 4: 10 500.

Amino Acids↗