A stable cell line inducibly expressing hERG1a/1b heteromeric channels

Ríos-Pérez, Erick Benjamín; Liu, Fang; Stevens-Sostre, Whitney A.; Eichel, Catherine A.; Silignavong, Jonathan; Robertson, Gail A.

doi:10.1016/j.vascn.2021.107081

Cited by 6 publications

(8 citation statements)

References 34 publications

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“…3 a,b). 100 μg/mL doxycycline significantly accelerated hERG current deactivation, demonstrating that the hERG 1b subunit was successfully co-expressed with hERG 1a 40 . Induced hERG 1b expression with doxycycline also depolarized the voltage dependence of deactivation without affecting the voltage dependence of activation (Fig.…”

Section: Resultsmentioning

confidence: 94%

“…These studies suggest that heteromeric hERG 1a/1b channels, comprising PAS-deficient hERG 1b subunits, would display accelerated VSD movement and reduced hysteresis compared to hERG 1a homomeric channels. To test this hypothesis, we repeated the experiments above using a cell line constitutively expressing hERG 1a, but with hERG 1b under control of an inducible promoter (see “ Methods ” 40 ) (Fig. 3 ).…”

Section: Resultsmentioning

confidence: 99%

“…We induced hERG 1b protein expression using 100 μg/mL doxycycline 48 h prior to recording. Details on the development and maintenance of this cell line were published previously 40 .…”

Section: Methodsmentioning

confidence: 99%

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Hysteretic hERG channel gating current recorded at physiological temperature

Jones

2022

Sci Rep

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Cardiac hERG channels comprise at least two subunits, hERG 1a and hERG 1b, and drive cardiac action potential repolarization. hERG 1a subunits contain a cytoplasmic PAS domain that is absent in hERG 1b. The hERG 1a PAS domain regulates voltage sensor domain (VSD) movement, but hERG VSD behavior and its regulation by the hERG 1a PAS domain have not been studied at physiological temperatures. We recorded gating charge from homomeric hERG 1a and heteromeric hERG 1a/1b channels at near physiological temperatures (36 ± 1 °C) using pulse durations comparable in length to the human ventricular action potential. The voltage dependence of deactivation was hyperpolarized relative to activation, reflecting VSD relaxation at positive potentials. These data suggest that relaxation (hysteresis) works to delay pore closure during repolarization. Interestingly, hERG 1a VSD deactivation displayed a double Boltzmann distribution, but hERG 1a/1b deactivation displayed a single Boltzmann. Disabling the hERG 1a PAS domain using a PAS-targeting antibody similarly transformed hERG 1a deactivation from a double to a single Boltzmann, highlighting the contribution of the PAS in regulating VSD movement. These data represent, to our knowledge, the first recordings of hERG gating charge at physiological temperature and demonstrate that VSD relaxation (hysteresis) is present in hERG channels at physiological temperature.

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Section: Resultsmentioning

confidence: 94%

Section: Resultsmentioning

confidence: 99%

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Hysteretic hERG channel gating current recorded at physiological temperature

Jones

2022

Sci Rep

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“…3a,b). 100 µg/ml doxycycline signi cantly accelerated hERG current deactivation, demonstrating that the hERG 1b subunit was successfully co-expressed with hERG 1a 39 . Induced hERG 1b expression with doxycycline also depolarized the voltage dependence of deactivation without affecting the voltage dependence of activation (Fig.…”

Section: Herg 1a Ionic Current Hysteresis At Physiological Temperaturementioning

confidence: 92%

“…We induced hERG 1b protein expression using 100 mg/ml doxycycline 48 hours prior to recording. Details on the development and maintenance of this cell line were published previously 39 .…”

Section: Materials Methodsmentioning

confidence: 99%

Hysteretic hERG Channel Gating Current Recorded At Physiological Temperature

Jones

2021

Preprint

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Cardiac hERG channels comprise at least two subunits, hERG 1a and hERG 1b, and drive cardiac action potential repolarization. hERG 1a subunits contain a cytoplasmic PAS domain that is absent in hERG 1b. The hERG 1a PAS domain regulates voltage sensor domain (VSD) movement, but hERG VSD behavior and its regulation by the hERG 1a PAS domain have not been studied at physiological temperatures. We recorded gating charge from homomeric hERG 1a and heteromeric hERG 1a/1b channels at near physiological temperatures (36 ± 1°C) using pulse durations comparable in length to the human ventricular action potential. The voltage dependence of deactivation was hyperpolarized relative to activation, reflecting VSD relaxation at positive potentials. These data suggest that relaxation (hysteresis) works to delay pore closure during repolarization. Interestingly, hERG 1a VSD deactivation displayed a double Boltzmann distribution, but hERG 1a/1b deactivation displayed a single Boltzmann. Disabling the hERG1a PAS domain using a PAS-targeting antibody similarly transformed hERG 1a deactivation from a double to a single Boltzmann, highlighting the contribution of the PAS in regulating VSD movement. These data represent, to our knowledge, the first recordings of hERG gating charge at physiological temperature and demonstrate that VSD relaxation (hysteresis) is present in hERG channels at physiological temperature.

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hERG agonists pose challenges to web-based machine learning methods for prediction of drug-hERG channel interaction

Harchi

Hancox

2023

Journal of Pharmacological and Toxicological Methods

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A stable cell line inducibly expressing hERG1a/1b heteromeric channels

Cited by 6 publications

References 34 publications

Hysteretic hERG channel gating current recorded at physiological temperature

Hysteretic hERG channel gating current recorded at physiological temperature

Hysteretic hERG Channel Gating Current Recorded At Physiological Temperature

hERG agonists pose challenges to web-based machine learning methods for prediction of drug-hERG channel interaction

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