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  • 1
    Digitale Medien
    Digitale Medien
    Springer
    The journal of membrane biology 112 (1989), S. 91-96 
    ISSN: 1432-1424
    Schlagwort(e): Paramecium ; patch clamp ; K channel ; Ca2+ dependence ; proteolysis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Summary The effects of proteolysis on a hyperpolarization- and Ca2+-dependent K channel from the surface membrane ofParamecium tetraurelia were examined in the inside-out excised patch mode. Treatment with trypsin, pronase or thermolysin removed the Ca2+-dependence of the channel activation, yielding an increase in channel activity greater than 2.5-fold at all Ca2+ concentrations between 10−4 and 10−8 m. Thermolysin addition-ally removed the voltage dependence of channel opening and gave the most activation among the three proteases tested. Proteolysis did not affect the single-channel conductance. In an analogy to the mechanism of activation of many Ca2+-dependent enzymes it is suggested that thisParamecium channel has a cytoplasmic inhibitory domain which can be removed by proteolysis, and that the physiological activation by Ca2+ is due to a temporary removal of this inhibition. Moreover, these findings indicate structural differences between depolarization-, Ca2+-dependent K channels (BK channels) and the hyperpolarization-, Ca2+-dependent K channels inParamecium.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    Springer
    The journal of membrane biology 112 (1989), S. 79-89 
    ISSN: 1432-1424
    Schlagwort(e): Paramecium ; patch clamp ; K channels ; Ca i 2+ -dependence ; hyperpolarization activated ; run-down
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Summary We have studied a class of Ca i 2+ -dependent K channels in inside-out excised membrane patches fromParamecium under patch clamp. Single channels had a conductance of 72 ±9.0 pS in a solution containing 100mM K+. The channels were selective for K+ over Rb+ with the permeability ratio of 1∶ 0.56. and over Na+, Cs+ or NH 4 + with a ratio 1∶〈0.1. The channel activity was dependent on Ca i 2+ , which was applied to the cytoplasmic side; the Ca i 2+ concentration for the half maximal activation was 2 μm. The Hill coefficient for the Ca i 2+ dependence of the channel activity was 2.58, indicating that more than two Ca i 2+ bindings are necessary for full activation. Unlike most Ca i 2+ -dependent K channels in other organisms, the channels inParamecium were slightly more active upon hyperpolarization than upon depolarization. The voltage dependence was fitted to a Boltzmann curve with 41.2 mV pere-fold change in channel activity. While a high Ca i 2+ concentration activated the channels, it also irreversibly reduced the channel activity over time. The decay of channel activity occurred faster at higher Ca i 2+ concentrations. Quaternary ammonium ions suppressed ion passage through the channel; more highly alkylated quaternary ammonium ions were more efficient in blocking. Ba i 2+ and Ca i 2+ were relatively ineffective in blockage. It was concluded that these Ca i 2+ -dependent K channels inParamecium are different from the previously described Ca i 2+ -dependent K channels, and are perhaps of a novel class.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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