Journal article
Blockade of Permeation by Potassium but Normal Gating of the G628S Nonconducting hERG Channel Mutant
Biophysical journal, Vol.101(3), pp.662-670
08/03/2011
DOI: 10.1016/j.bpj.2011.06.028
PMCID: PMC3145283
PMID: 21806934
Abstract
G628S is a mutation in the signature sequence that forms the selectivity filter of the human ether-a-go-go-related gene (hERG) channel (GFG) and is associated with long-QT2 syndrome. G628S channels are known to have a dominant-negative effect on hERG currents, and the mutant is therefore thought to be nonfunctional. This study aims to assess the physiological mechanism that prevents the surface-expressing G628S channels from conducting ions. We used voltage-clamp fluorimetry along with two-microelectrode voltage clamping in
Xenopus
oocytes to confirm that the channels express well at the surface, and to show that they are actually functional, with activation kinetics comparable to that of wild-type, and that the mutation leads to a reduced selectivity to potassium. Although ionic currents are not detected in physiological solutions, removing extracellular K
+
results in the appearance of an inward Na
+
-dependent current. Using whole-cell patch clamp in mammalian transfected cells, we demonstrate that the G628S channels conduct Na
+
, but that this can be blocked by both intracellular and higher-than-physiological extracellular K
+
. Using solutions devoid of K
+
allows the appearance of nA-sized Na
+
currents with activation and inactivation gating analogous to wild-type channels. The G628S channels are functionally conducting but are normally blocked by intracellular K
+
.
Details
- Title: Subtitle
- Blockade of Permeation by Potassium but Normal Gating of the G628S Nonconducting hERG Channel Mutant
- Creators
- Zeineb Es-Salah-Lamoureux - Department of Anesthesiology, Pharmacology, and Therapeutics, University of British Columbia, Vancouver, British Columbia, CanadaPing Yu Xiong - Department of Anesthesiology, Pharmacology, and Therapeutics, University of British Columbia, Vancouver, British Columbia, CanadaSamuel J Goodchild - Department of Anesthesiology, Pharmacology, and Therapeutics, University of British Columbia, Vancouver, British Columbia, CanadaChristopher A Ahern - Department of Anesthesiology, Pharmacology, and Therapeutics, University of British Columbia, Vancouver, British Columbia, CanadaDavid Fedida - Department of Anesthesiology, Pharmacology, and Therapeutics, University of British Columbia, Vancouver, British Columbia, Canada
- Resource Type
- Journal article
- Publication Details
- Biophysical journal, Vol.101(3), pp.662-670
- Publisher
- The Biophysical Society
- DOI
- 10.1016/j.bpj.2011.06.028
- PMID
- 21806934
- PMCID
- PMC3145283
- ISSN
- 0006-3495
- eISSN
- 1542-0086
- Language
- English
- Date published
- 08/03/2011
- Academic Unit
- Molecular Physiology and Biophysics; Iowa Neuroscience Institute
- Record Identifier
- 9984070729602771
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