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Related Experiment Videos

Anion permeation in Ca(2+)-activated Cl(-) channels.

Z Qu1, H C Hartzell

  • 1Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.

The Journal of General Physiology
|December 1, 2000
PubMed
Summary

Calcium(2+)-activated chloride channels (Cl(Ca)Cs) from Xenopus oocytes show distinct anion permeation properties. These channels exhibit voltage-dependent block and pH sensitivity, offering insights into pore characteristics and differing from other chloride channels.

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Area of Science:

  • Molecular Biology
  • Ion Channel Physiology
  • Biophysics

Background:

  • Calcium(2+)-activated chloride channels (Cl(Ca)Cs) are crucial for cellular processes, responding to elevated intracellular calcium.
  • Understanding their ion permeation mechanisms is key to elucidating their physiological roles.

Purpose of the Study:

  • To investigate the biophysical properties of anion permeation through Cl(Ca)Cs in Xenopus oocytes.
  • To characterize the pore diameter, ion selectivity, and voltage-dependent block of Cl(Ca)Cs.
  • To explore the influence of pH and permeant anions on channel function and calcium affinity.

Main Methods:

  • Utilized excised inside-out and outside-out membrane patches from Xenopus oocytes.
  • Performed electrophysiological recordings to measure ion permeation and channel block.

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  • Analyzed current-voltage relationships and employed bi-ionic conditions to determine relative ion permeabilities and affinities.

    • Main Results:

    • Cl(Ca)Cs demonstrated moderate chloride selectivity over sodium (P(Na)/P(Cl) = 0.1) and a low apparent affinity for chloride (Kd = 73 mM).
    • The pore diameter was estimated to be greater than 0.6 nm, with a specific sequence of anion relative permeabilities and conductances observed.
    • Permeant anions caused voltage-dependent block, and channel affinity for calcium increased with anion occupancy. pH significantly affected channel current, indicating potential proton titration or hydroxyl ion block.

    Conclusions:

    • Anion selectivity is influenced by hydration energy but not solely determined by it.
    • Cl(Ca)Cs may function as multi-ion pores with weak ion interactions.
    • The distinct permeation properties of Cl(Ca)Cs differentiate them from CFTR and ClC-1 channels, providing novel insights into their pore structure and function.