Constitutive opening of the Kv7.2 pore activation gate causes KCNQ2- developmental encephalopathy

• 成果类型: Article
• 署名作者: Nappi, Mario; Alberini, Giulio; Berselli, Alessandro; Roscioni, Agnese; Soldovieri, Maria Virginia; Servettini, Ilenio; Barrese, Vincenzo; Weckhuysen, Sarah; Chiu, Ting-Gee Annie; Scheffer, Ingrid E.; Benfenati, Fabio; Maragliano, Luca; Miceli, Francesco; Taglialatela, Maurizio
• 署名单位: University of Naples Federico II; Istituto Italiano di Tecnologia - IIT; Marche Polytechnic University; University of Molise; Flanders Institute for Biotechnology (VIB); University of Antwerp; University of Antwerp; University of Antwerp; Florey Institute of Neuroscience & Mental Health; Howard Florey Institute Affiliates; University of Melbourne; University of Melbourne; Florey Institute of Neuroscience & Mental Health; University of Melbourne; Murdoch Children's Research Institute; Royal Children's Hospital Melbourne
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-13634
• DOI: 10.1073/pnas.2412388121
• 发表日期: 2024-12-03
• 关键词: onset epileptic encephalopathy; genotype-phenotype correlations; molecular-dynamics simulations; voltage sensor; potassium channels; force-field; mutations; epilepsies; k(v)7.2; validation

摘要

Pathogenic variants in KCNQ2 encoding Kv7.2 voltage- gated potassium channel subunits cause developmental encephalopathies ( KCNQ2- encephalopathies), both with and without epilepsy. We herein describe the clinical, in vitro, and in silico features of two encephalopathy- causing variants (A317T, L318V) in Kv7.2 affecting two consecutive residues in the S6 activation gate that undergoes large structural rearrangements during pore opening; the disease- causing A356T variant in KCNQ3, paralogous to the A317T variant in KCNQ2, was also investigated. Currents through KCNQ2 mutant channels displayed increased density, hyperpolarizing shifts in activation gating, faster activation and slower deactivation kinetics, and resistance to changes in the cellular concentrations ofphosphatidylinositol 4,5- bisphosphate (PIP2), a critical regulator of Kv7 channel function; all these features are consistent with a strong gain- of- function effect. An increase in the probability of single- channel opening, with no change in membrane abundance or single- channel conductance, was responsible for the observed gain- of- function effects. All- atom molecular dynamics simulations revealed that the mutations widened the inner pore gate and stabilized a constitutively open channel configuration in the closed state, with minimal effects on the open conformation. Thus, mutation- induced stabilization of the inner pore gate open configuration is a molecular pathogenetic mechanism for KCNQ2- related encephalopathies.