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  • Key words Metalloenzymes  (1)
  • cell nuclei fusion  (1)
  • Springer  (2)
  • American Chemical Society
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  • Springer  (2)
  • American Chemical Society
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  • 1
    ISSN: 1432-1424
    Keywords: Cell membrane fusion ; cell nuclei fusion ; cell culture ; MDCK cells ; lectins ; acridine orange
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The evaluation of the intracellular signal train and its regulatory function in controlling transepithelial transport with electrophysiological methods often requires intracellular measurements with microelectrodes. However, multiple impalements in epithelial cells are hampered by the small size of the cells. In an attempt to avoid these problems we fused cells of an established cell line, Madin Darby canine kidney cells, originally derived from dog kidney, to “giant” cells by applying a modified polyethylene glycol method. During trypsin-induced detachment from the ground of the petri dish, individual cells grown in a monolayer incorporate volume and mainly lose basolateral plasma membrane by extrusion. By isovolumetric cell-to-cell fusion, spherical “giant” cells are formed within 2 hr. During this process a major part of the individual cell plasma membranes is internalized. Over three weeks following cell plasma membrane fusion degradation of single cell nuclei and cell nuclear fusion occurs. We conclude that this experimental approach opens the possibility to investigate ion transport of epithelia in culture by somatic cell genetic techniques.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1327
    Keywords: Key words Metalloenzymes ; Nuclear magnetic resonance ; Aromatic compounds ; biodegradation ; hydration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract  Water proton T 1 –1 measurements at magnetic fields between 0.01 and 50 MHz [nuclear magnetic relaxation dispersion (NMRD) measurements] have been performed on solutions of phthalate dioxygenase (PDO) reconstituted at the catalytic iron site with copper(II) or manganese(II). The data show evidence of a weakly coordinated water molecule in CuPDO; in the presence of the substrate, phthalate, this water appears to become even less tightly bound, and an additional tightly coordinated water can be detected. In PDO reconstituted with manganese, one tightly coordinated water is detected in the presence and in the absence of phthalate. An attempt is made to reconcile these data with low-temperature near-IR magnetic circular dichroism and X-ray absorption data, which show that PDO reconstituted with iron or cobalt is six-coordinate in the absence of substrate and five-coordinate in the presence of substrate.
    Type of Medium: Electronic Resource
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