Class III antiarrhythmic drugs block the rapid delayed rectifier K+ channel hERG and prolong ventricular repolarization, measured on the ECG as a lengthened QT interval. Drug-induced hERG blockade and QT prolongation also occurs as an unintended side effect of some common medications and increases the risk of torsades de pointes, a ventricular arrhythmia that can degenerate into ventricular fibrillation and sudden death. An understanding of the molecular determinants of drug binding to hERG channels would facilitate design of new drugs devoid of this dangerous side-effect. Treatment of congenital or drug-induced long QT syndrome is presently inadequate but a recently discovered hERG channel agonist represents a potential new therapy. The goals of this project are to characterize the mechanisms of action, and the structural basis of the binding site of hERG channel blockers and activators. We previously used an Ala-scanning mutagenesis approach to define the major structural determinants of the hERG channel that can account for block by high affinity blockers such as cisapride and terfenadine. Two aromatic residues in the S6 domain (Tyr652, Phe656) and three residues located at the base of the pore helix (Thr623, Ser624 and Val625) were required for potent block by structurally diverse drugs.
In Aim 1, we will refine the molecular determinants for hERG block by high affinity ligands.
Aim 2 will define the binding site for low affinity ligands (antibiotics) using site-directed mutagenesis and voltage clamp of mutant channels expressed in Xenopus oocytes.
In Aim 3, we will determine the binding site and molecular basis of altered gating induced by two hERG channel activators, a fenamate and a novel quinolinylpiperidine.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL055236-14
Application #
7743379
Study Section
Electrical Signaling, Ion Transport, and Arrhythmias Study Section (ESTA)
Program Officer
Lathrop, David A
Project Start
1996-07-01
Project End
2011-07-31
Budget Start
2009-12-01
Budget End
2011-07-31
Support Year
14
Fiscal Year
2010
Total Cost
$362,911
Indirect Cost
Name
University of Utah
Department
Physiology
Type
Schools of Medicine
DUNS #
009095365
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Gardner, Alison; Wu, Wei; Thomson, Steven et al. (2017) Molecular Basis of Altered hERG1 Channel Gating Induced by Ginsenoside Rg3. Mol Pharmacol 92:437-450
Wu, Wei; Gardner, Alison; Sachse, Frank B et al. (2016) Ginsenoside Rg3, a Gating Modifier of EAG Family K+ Channels. Mol Pharmacol 90:469-82
Wu, Wei; Sanguinetti, Michael C (2016) Molecular Basis of Cardiac Delayed Rectifier Potassium Channel Function and Pharmacology. Card Electrophysiol Clin 8:275-84
Gardner, Alison; Sanguinetti, Michael C (2015) C-Linker Accounts for Differential Sensitivity of ERG1 and ERG2 K+ Channels to RPR260243-Induced Slow Deactivation. Mol Pharmacol 88:19-28
Wu, Wei; Gardner, Alison; Sanguinetti, Michael C (2015) The Link between Inactivation and High-Affinity Block of hERG1 Channels. Mol Pharmacol 87:1042-50
Wu, Wei; Gardner, Alison; Sanguinetti, Michael C (2015) Concatenated hERG1 tetramers reveal stoichiometry of altered channel gating by RPR-260243. Mol Pharmacol 87:401-9
Thomson, Steven J; Hansen, Angela; Sanguinetti, Michael C (2014) Concerted all-or-none subunit interactions mediate slow deactivation of human ether-à-go-go-related gene K+ channels. J Biol Chem 289:23428-36
Wu, Wei; Gardner, Alison; Sanguinetti, Michael C (2014) Cooperative subunit interactions mediate fast C-type inactivation of hERG1 K+ channels. J Physiol 592:4465-80
Sanguinetti, Michael C (2014) HERG1 channel agonists and cardiac arrhythmia. Curr Opin Pharmacol 15:22-7
Wu, Wei; Sachse, Frank B; Gardner, Alison et al. (2014) Stoichiometry of altered hERG1 channel gating by small molecule activators. J Gen Physiol 143:499-512

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