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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 80 (1984), S. 175-183 
    ISSN: 1432-1424
    Keywords: Chara ; electrogenic pump ; chord conductance ; I–V curve ; kinetic model ; computer simulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The current-voltage curve of theChara membrane was obtained by applying a slow ramp depo- and hyperpolarization by use of voltage clamp. With the progress of poisoning by DCCD (dicyclohexylcarbodiimide) theI–V curve moved by about 50 mV (depolarization) along the voltage axis, reducing its slope, and finally converged to thei d -V curve of the passive diffusion channel. Changes ofi p -V curve of the electrogenic pump channel could be obtained by subtracting the latter from the former. The sigmoidali p -V curve could be simulated satisfactorily by adopting a simple reaction kinetic model. Kinetic parameters of the successive changes of state of the H+ ATPase could be evaluated. Changes of these kinetic parameters during inhibition gave useful information about the molecular mechanism of the electrogenic pump. Depolarization of the membrane potential, decrease of membrane conductance, and decrease of pump current during inhibition of the pump with DCCD are caused mainly by the decrease of conductance of the pump channel. The decrease of this pump conductance is caused principally by a marked decrease of the rate constant for releasing H+ to the outside.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1424
    Keywords: Chara ; electrogenic pump ; chord conductance ; I–V curve ; kinetic model ; pH dependence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The current-voltage curve of theChara membrane was obtained by applying a slow ramp de- and hyperpolarization by use of voltage clamp. By inhibiting the electrogenic pump with 50μm DCCD (dicyclohexylcarbodiimide), theI–V curve approached a steady state within 100 min, which gave thei d -V curve of the passive diffusion channel. Thei p -V curve of the electrogenic pump channel was obtained by subtracting the latter from the former. With the increase of external pH, thei d -V curve showed only a slight change, while thei p -V curve of the pump channel showed almost a parallel shift, in the hyperpolarizing direction, along the voltage axis in the pH range between 6.5 and 7.5. The sigmoidali p -V curve in this pH range could be simulated satisfactorily with the five-state model reported previously (U. Kishimoto, N. Kami-ike, Y. Takeuchi & T. Ohkawa,J. Membrane Biol. 80:175–183, 1984) as well as with a lumped two-state model presented in this report. The analysis based on these models suggests that the electrogenic pump of theChara membrane is mainly a 2H+/1ATP pump. The forward rate constant in the voltage-dependent step increased with the increase of external pH, while the backward one decreased. On the other hand, the forward rate constant in the voltage-independent step remained almost unchanged with the increase of external pH, while the backward one increased markedly. The pump conductance at the resting membrane potential showed either a slight increase or a decrease with the increase of external pH, depending on the sample. Nevertheless, the pump current showed generally a slight increase with the increase of external pH.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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