Novel Kv7.1-phosphatidylinositol 4,5-bisphosphate interaction sites uncovered by charge neutralization scanning

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Novel Kv7.1-phosphatidylinositol 4,5-bisphosphate interaction sites uncovered by charge neutralization scanning. / Eckey, Karina; Wrobel, Eva; Strutz-Seebohm, Nathalie; Pott, Lutz; Schmitt, Nicole; Seebohm, Guiscard.

In: The Journal of Biological Chemistry, Vol. 289, No. 33, 15.08.2014, p. 22749-58.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Eckey, K, Wrobel, E, Strutz-Seebohm, N, Pott, L, Schmitt, N & Seebohm, G 2014, 'Novel Kv7.1-phosphatidylinositol 4,5-bisphosphate interaction sites uncovered by charge neutralization scanning', The Journal of Biological Chemistry, vol. 289, no. 33, pp. 22749-58. https://doi.org/10.1074/jbc.M114.589796

APA

Eckey, K., Wrobel, E., Strutz-Seebohm, N., Pott, L., Schmitt, N., & Seebohm, G. (2014). Novel Kv7.1-phosphatidylinositol 4,5-bisphosphate interaction sites uncovered by charge neutralization scanning. The Journal of Biological Chemistry, 289(33), 22749-58. https://doi.org/10.1074/jbc.M114.589796

Vancouver

Eckey K, Wrobel E, Strutz-Seebohm N, Pott L, Schmitt N, Seebohm G. Novel Kv7.1-phosphatidylinositol 4,5-bisphosphate interaction sites uncovered by charge neutralization scanning. The Journal of Biological Chemistry. 2014 Aug 15;289(33):22749-58. https://doi.org/10.1074/jbc.M114.589796

Author

Eckey, Karina ; Wrobel, Eva ; Strutz-Seebohm, Nathalie ; Pott, Lutz ; Schmitt, Nicole ; Seebohm, Guiscard. / Novel Kv7.1-phosphatidylinositol 4,5-bisphosphate interaction sites uncovered by charge neutralization scanning. In: The Journal of Biological Chemistry. 2014 ; Vol. 289, No. 33. pp. 22749-58.

Bibtex

@article{e7327b2f92f74b1b9bfbf84f9fb6cb4e,
title = "Novel Kv7.1-phosphatidylinositol 4,5-bisphosphate interaction sites uncovered by charge neutralization scanning",
abstract = "Kv7.1 to Kv7.5 α-subunits belong to the family of voltage-gated potassium channels (Kv). Assembled with the β-subunit KCNE1, Kv7.1 conducts the slowly activating potassium current IKs, which is one of the major currents underlying repolarization of the cardiac action potential. A known regulator of Kv7 channels is the lipid phosphatidylinositol 4,5-bisphosphate (PIP2). PIP2 increases the macroscopic current amplitude by stabilizing the open conformation of 7.1/KCNE1 channels. However, knowledge about the exact nature of the interaction is incomplete. The aim of this study was the identification of the amino acids responsible for the interaction between Kv7.1 and PIP2. We generated 13 charge neutralizing point mutations at the intracellular membrane border and characterized them electrophysiologically in complex with KCNE1 under the influence of diC8-PIP2. Electrophysiological analysis of corresponding long QT syndrome mutants suggested impaired PIP2 regulation as the cause for channel dysfunction. To clarify the underlying structural mechanism of PIP2 binding, molecular dynamics simulations of Kv7.1/KCNE1 complexes containing two PIP2 molecules in each subunit at specific sites were performed. Here, we identified a subset of nine residues participating in the interaction of PIP2 and Kv7.1/KCNE1. These residues may form at least two binding pockets per subunit, leading to the stabilization of channel conformations upon PIP2 binding.",
keywords = "Action Potentials, Amino Acid Substitution, Animals, Binding Sites, Humans, KCNQ1 Potassium Channel, Phosphatidylinositol 4,5-Diphosphate, Point Mutation, Protein Binding, Xenopus laevis",
author = "Karina Eckey and Eva Wrobel and Nathalie Strutz-Seebohm and Lutz Pott and Nicole Schmitt and Guiscard Seebohm",
note = "{\circledC} 2014 by The American Society for Biochemistry and Molecular Biology, Inc.",
year = "2014",
month = "8",
day = "15",
doi = "10.1074/jbc.M114.589796",
language = "English",
volume = "289",
pages = "22749--58",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "33",

}

RIS

TY - JOUR

T1 - Novel Kv7.1-phosphatidylinositol 4,5-bisphosphate interaction sites uncovered by charge neutralization scanning

AU - Eckey, Karina

AU - Wrobel, Eva

AU - Strutz-Seebohm, Nathalie

AU - Pott, Lutz

AU - Schmitt, Nicole

AU - Seebohm, Guiscard

N1 - © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

PY - 2014/8/15

Y1 - 2014/8/15

N2 - Kv7.1 to Kv7.5 α-subunits belong to the family of voltage-gated potassium channels (Kv). Assembled with the β-subunit KCNE1, Kv7.1 conducts the slowly activating potassium current IKs, which is one of the major currents underlying repolarization of the cardiac action potential. A known regulator of Kv7 channels is the lipid phosphatidylinositol 4,5-bisphosphate (PIP2). PIP2 increases the macroscopic current amplitude by stabilizing the open conformation of 7.1/KCNE1 channels. However, knowledge about the exact nature of the interaction is incomplete. The aim of this study was the identification of the amino acids responsible for the interaction between Kv7.1 and PIP2. We generated 13 charge neutralizing point mutations at the intracellular membrane border and characterized them electrophysiologically in complex with KCNE1 under the influence of diC8-PIP2. Electrophysiological analysis of corresponding long QT syndrome mutants suggested impaired PIP2 regulation as the cause for channel dysfunction. To clarify the underlying structural mechanism of PIP2 binding, molecular dynamics simulations of Kv7.1/KCNE1 complexes containing two PIP2 molecules in each subunit at specific sites were performed. Here, we identified a subset of nine residues participating in the interaction of PIP2 and Kv7.1/KCNE1. These residues may form at least two binding pockets per subunit, leading to the stabilization of channel conformations upon PIP2 binding.

AB - Kv7.1 to Kv7.5 α-subunits belong to the family of voltage-gated potassium channels (Kv). Assembled with the β-subunit KCNE1, Kv7.1 conducts the slowly activating potassium current IKs, which is one of the major currents underlying repolarization of the cardiac action potential. A known regulator of Kv7 channels is the lipid phosphatidylinositol 4,5-bisphosphate (PIP2). PIP2 increases the macroscopic current amplitude by stabilizing the open conformation of 7.1/KCNE1 channels. However, knowledge about the exact nature of the interaction is incomplete. The aim of this study was the identification of the amino acids responsible for the interaction between Kv7.1 and PIP2. We generated 13 charge neutralizing point mutations at the intracellular membrane border and characterized them electrophysiologically in complex with KCNE1 under the influence of diC8-PIP2. Electrophysiological analysis of corresponding long QT syndrome mutants suggested impaired PIP2 regulation as the cause for channel dysfunction. To clarify the underlying structural mechanism of PIP2 binding, molecular dynamics simulations of Kv7.1/KCNE1 complexes containing two PIP2 molecules in each subunit at specific sites were performed. Here, we identified a subset of nine residues participating in the interaction of PIP2 and Kv7.1/KCNE1. These residues may form at least two binding pockets per subunit, leading to the stabilization of channel conformations upon PIP2 binding.

KW - Action Potentials

KW - Amino Acid Substitution

KW - Animals

KW - Binding Sites

KW - Humans

KW - KCNQ1 Potassium Channel

KW - Phosphatidylinositol 4,5-Diphosphate

KW - Point Mutation

KW - Protein Binding

KW - Xenopus laevis

U2 - 10.1074/jbc.M114.589796

DO - 10.1074/jbc.M114.589796

M3 - Journal article

C2 - 24947509

VL - 289

SP - 22749

EP - 22758

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 33

ER -

ID: 130330216