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  • 1
    ISSN: 1432-2145
    Keywords: Key words Profilin ; Tobacco pollen ; Nicotiana tabacum ; cDNA cloning ; Isoforms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Profilins are actin-binding proteins in eukaryotes which participate in the phosphoinositide pathway via binding to PIP2. Using polyclonal rabbit sera raised against plant profilins, the occurrence of several profilin isoforms is demonstrated in two-dimensionally analyzed tobacco pollen extracts. The cDNAs coding for two novel tobacco profilin isoforms (ntPro2, ntPro3) were isolated from a pollen cDNA library by antibody screening. When the cDNA and deduced amino acid sequences of the two isoforms were compared with a previously isolated tobacco pollen profilin cDNA (ntPro1), significant differences were noted in the non-coding regions, whereas the coding sequences, in particular the functional domains, showed little variation. The cDNAs coding for the three tobacco profilin isoforms were expressed in Escherichia coli and shown to bind comparably to different anti-profilin antisera. The high degree of similarity among the different tobacco pollen profilin isoforms points to functional equivalence. Assuming that the presence of profilin is indispensable to the control of the large amounts of actin present in pollen, the occurrence of different profilin isoforms in pollen is interpreted to represent a protective mechanism against loss of profilin functions.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1615-6102
    Keywords: Actin-binding protein ; Green algae ; Immunolocalization ; Micrasterias denticulata ; Profilin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Profilin is detected by means of immunoblotting in the green algaMicrasterias denticulata at a molecular mass of about 14 kDa with antibodies against celery root profilin, recombinant tobacco profilin, and recombinant birch profilin. Poly-L-proline purification ofM. denticulata extracts leads to a single band at 14 kDa. By means of immunoelectron microscopy first evidence is provided for the presence of profilin in a microtubule center associated with the migrating nucleus in high-pressure-frozen and freeze-substituted cells. Colocalization with a particular filamen-tous-actin aggregation in the same area suggests a role of profilin in the process of nuclear migration. Moreover staining is visible in several areas of the nucleus including the nucleolus, heterochromatin, and nuclear-pore complexes. An even distribution of profilin is found throughout the cytoplasm at the confocal-microscopy level as well as by immunogold labeling. Occasionally also areas at the surface of the chloroplast are stained.
    Type of Medium: Electronic Resource
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