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  • Glycoproteins  (2)
  • Mucor rouxii  (2)
  • chitosomes  (2)
  • 1
    ISSN: 1572-9699
    Keywords: Chitin synthetase ; chitosomes ; Mucorales
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Stability of chitin synthetase in cell-free extracts from mycelial fungi was markedly improved by the presence of sucrose in the homogenization media. Breakage of mycelium in sucrose-containing buffer yielded enzyme preparations from which chitosomal chitin synthetase could be purified by a procedure involving ammonium sulfate precipitation, gel filtration and centrifugation in sucrose density gradients. Purified chitosomes catalyzed the synthesis of chitin microfibrils in vitro upon incubation with substrate and activators. Chitosomal chitin synthetase from the filamentous form of M. rouxii was similar to the enzyme from yeast cells, except for the poorer stability and diminished sensitivity to GlcNAc activation of the former.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 122 (1984), S. 178-190 
    ISSN: 1615-6102
    Keywords: Chitin synthetase localization ; Chitosome sedimentation ; Fungus ; Miniorganelles ; Mucor rouxii ; Plasma membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This study was undertaken to assess the distribution and localization of chitin synthetase in a fungal cell and to evaluate the sedimentation behavior of chitosomes (microvesicular containers of chitin synthetase). Chitosomes were isolated from cell-free extracts of yeast cells ofMucor rouxii by rate-zonal and isopycnic sedimentation in sucrose density gradients. Because of their small size and low density, chitosomes were effectively separated from other subcellular particles. Rate-zonal sedimentation was a suitable final step for isolating chitosomes as long as ribosomes had been eliminated by enzymic digestion. By isopycnic centrifugation, chitosomes could be separated directly from a crude cell-free extract; they cosedimented with a sharp symmetrical peak of chitin synthetase at a buoyant density of d=1.14–1.15g/cm3; the only significant contaminants were particles of fatty acid synthetase complex. From such sedimentations, we estimated that 80–85% of the chitin synthetase activity in the cell-free extract was associated with chitosomes; the rest was found in two smaller peaks sedimenting at d=1.19–1.20 and d=1.21–1.22 (5–10%), and in the cell wall fraction (5–10%). By consecutive rate-zonal and isopycnic sedimentations, chitosome preparations with relatively few contaminating particles were obtained. Potassium/sodium phosphate buffer (pH 6.5)+MgCl2 was the most effective isolation medium for chitosomes. Other buffers such as TRIS-MES+MgCl2 led to massive aggregation of chitosomes and a change in sedimentation properties. This tendency of chitosomes to aggregate could explain why most of the chitin synthetase activity of a fungus is sometimes found associated with other subcellular structures,e.g., plasma membrane.
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  • 3
    ISSN: 0147-5975
    Keywords: Phycomyces ; chitin microfibrils ; chitin synthesis ; chitosomes ; sporangiophores
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Experimental Mycology 3 (1979), S. 351-362 
    ISSN: 0147-5975
    Keywords: Mannan ; Mucor rouxii ; cell wall ; fungal growth ; fungus ; mannosyl transferase ; morphogenesis
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-072X
    Keywords: Neurospora crassa ; “Slime” variant ; Cell coat ; Exocellular proteins ; Glycoproteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of the “slime” strain of Neurospora crassa synthesize a coherent extracellular material which remains attached to the cell surface, but is released into the liquid medium by shaking. The material was purified and studied by different criteria. By electron microscopy it appears as long wavy sheets which strongly bind concanavalin A, but not wheat germ agglutinin, and maintain their integrity in the absence of structural polysaccharides. Analysis of the purified material revealed that it was free of contaminating membranes; it contained more than 70% protein, 1% neutral sugars (glucose, mannose, fucose and galactose), less than 2% lipids and ca. 4% not-characterized hexosaminelike compounds. Its polypeptide pattern as determined by PAGE was complex. The significance of this material is discussed.
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  • 6
    ISSN: 1432-072X
    Keywords: Neurospora crassa ‘Slime” variant ; Cell coat ; Glycoproteins ; Self-assembling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The proteinaceous extracellular material (PEM) synthesized by the cells of the ‘slime” strain of Neurospora crassa (see Martinez et al. 1989) was solubilized by treatment with urea or guanidine. Removal of these chemicals by dialysis, caused reassembly of the solubilized proteins into material with the same microscopic appearance as the original PEM. Polypeptide patterns from both native and reassembled structures were identical. Dialysis-mediated reassembly of the solubilized proteins appeared to be dependent on both concentration of the soluble macromolecules and time. Gel chromatography of PEM solubilized with different agents revealed two discrete populations of complexes with molecular masses of 1,500 and 500 kDa respectively. These were able to reassemble into lamellar structures with a variable degree of efficiency.
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