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  • 1
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 671 (1992), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The potassium-translocating Kdp-ATPase of Escherichia coli shares common functional properties with eukaryotic P-type ATPases. The KdpB subunit has been identified as the catalytic subunit forming the phosphorylated intermediate. Substitution of Asp 307 in KdpB by Glu, Asn, Gin, Tyr, His, Ala or Ser by site-directed mutagenesis and the subsequent transfer of the point mutations to the chromosome revealed that the mutants were not functioning with respect to cell growth at low K’ concentrations and ATPase activity as well as phosphorylation capacity of the purified Kdp complex. These findings indicate that Asp-307 in KdpB is the phosphorylation site of the Kdp-ATPase. In contrast, replacement of the close but non-conserved Asp-300 by Asn or Glu has no immediate influence on the enzyme functions tested. However, the Km for K+ of the ATPase activity has been increased 30-fold compared with the wild-type enzyme.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-072X
    Keywords: Potassium transport ; High-affinity transport system ; Kdp-ike potassium ATPase ; Expression ; Immunological cross-reactivity ; Internal pH regulation ; Photosynthetic bacteria (Rhodobacter sphaeroides, Rhodobacter capsulatus and Rhodospirillum rubrum)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of the purple non-sulphur bacterium Rhodobacter sphaeroides express a high-affinity K+ uptake system when grown in media with low K+ concentrations. Antibodies againts the catalytic KdpB protein or the whole KdpABC complex of Escherichia coli crossreact with a 70.0 kDa R. sphaeroides protein that was expressed only in cells grown in media with low K+ concentrations. In membranes derived from R. sphaeroides cells grown with low K+ concentrations (induced cells), a high ATPase activity could be detected when assayed in Tris-HCl pH 8.0 containing 1 mM MgSO4. This ATPase activity increased upon addition of 1 mM KCl from 166 to 289 μmol ATP hydrolysed x min-1 x g protein-1 (1.7-fold stimulation). The K+-stimulated ATPase activity was inhibited approximately 93% by 0.5 mM vanadate but hardly by N,N′-dicyclohexylcarbo-diimide (DCCD). These results indicate that the inducible K+-ATPase in R. sphaeroides resembles the Kdp K+-translocating ATPase of Escherichia coli. This Kdp-like transport system is also expressed in R. capsulatus and Rhodospirillum rubrum during growth in media with low K+ concentrations suggesting a wide distribution of this transport system among phototrophic bacteria.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1831
    Keywords: Key wordsBorrelia burgdorferi ; Lyme borreliosis ; Cloning ; Pathogenic potential ; Antigenic variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To study clonal polymorphism of Borrelia burgdorferi antigens in the course of an experimental infection sequence, the low-passage tick isolate ZS7 was cloned by two rounds of agar subsurface plating. The resulting clones showed a variable pathogenic potential after experimental infection of C.B-17.scid mice. The test clone 4.2.II, selected for virulence by two passages in immunodeficient scid mice, failed to establish a successful infection in immunocompetent AKR/N mice, indicating the loss of pathogenicity traits required for evasion of the specific immune response. Cloning of natural or clinical B. burgdorferi isolates is a prerequisite for analyzing genetic and antigenic variation of the pathogen. However, the inevitable propagation in artificial media during cloning may lead to a loss of pathogenic features rendering the subsequent experimental infection of animals impossible. A combined procedure of in vitro cloning and in vivo selection also does not solve the dilemma because B. burgdorferi variants arise by recombinatorial processes in the pathogen's dynamic genome during the course of infection. Consequently, the resulting bacterial isolates from infected animal tissues represent again non-clonal, heterogeneous B. burgdorferi populations. In principle, cloning of a B. burgdorferi population is the appropriate method to analyze the polymorphism of individual molecules during infection. As a caveat, however, one has to envisage that during propagation of individual clones in vitro and in vivo independent genetic variations (epigenetic or mutational) may occur with consequences on the virulence of the clones. This may complicate the delineation of a clear correlation between the antigenic polymorphism observed and the change of virulence.
    Type of Medium: Electronic Resource
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