Ion permeation of AQP6 water channel protein. Single channel recordings after Hg2+ activation.

Akihiro Hazama, David Kozono, William B. Guggino, Peter Agre, Masato Yasui

Research output: Contribution to journalArticle

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Abstract

Aquaporin-6 (AQP6) has recently been identified as an intracellular vesicle water channel with anion permeability that is activated by low pH or HgCl2. Here we present direct evidence of AQP6 channel gating using patch clamp techniques. Cell-attached patch recordings of AQP6 expressed in Xenopus laevis oocytes indicated that AQP6 is a gated channel with intermediate conductance (49 picosiemens in 100 mm NaCl) induced by 10 microm HgCl2. Current-voltage relationships were linear, and open probability was fairly constant at any given voltage, indicating that Hg2+-induced AQP6 conductance is voltage-independent. The excised outside-out patch recording revealed rapid activation of AQP6 channels immediately after application of 10 microm HgCl2. Reduction of both Na+ and Cl- concentrations from 100 to 30 mm did not shift the reversal potential of the Hg2+-induced AQP6 current, suggesting that Na+ is as permeable as Cl-. The Na+ permeability of Hg2+-induced AQP6 current was further demonstrated by 22Na+ influx measurements. Site-directed mutagenesis identified Cys-155 and Cys-190 residues as the sites of Hg2+ activation both for water permeability and ion conductance. The Hill coefficient from the concentration-response curve for Hg2+-induced conductance was 1.1 +/- 0.3. These data provide the first evidence of AQP6 channel gating at a single-channel level and suggest that each monomer contains the pore region for ions based on the number of Hg2+-binding sites and the kinetics of Hg2+-activation of the channel.

Original languageEnglish
Pages (from-to)29224-29230
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number32
Publication statusPublished - 2002 Aug 9
Externally publishedYes

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Aquaporin 6
Aquaporins
Permeation
Chemical activation
Ions
Mercuric Chloride
Permeability
Electric potential
Mutagenesis
Clamping devices
Xenopus laevis
Patch-Clamp Techniques
Site-Directed Mutagenesis
Oocytes
Anions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Ion permeation of AQP6 water channel protein. Single channel recordings after Hg2+ activation. / Hazama, Akihiro; Kozono, David; Guggino, William B.; Agre, Peter; Yasui, Masato.

In: Journal of Biological Chemistry, Vol. 277, No. 32, 09.08.2002, p. 29224-29230.

Research output: Contribution to journalArticle

Hazama, Akihiro ; Kozono, David ; Guggino, William B. ; Agre, Peter ; Yasui, Masato. / Ion permeation of AQP6 water channel protein. Single channel recordings after Hg2+ activation. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 32. pp. 29224-29230.
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