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The connexin26 human mutation N14K disrupts cytosolic intersubunit interactions and promotes channel opening
A group of human mutations within the N-terminal (NT) domain of connexin 26 (Cx26) hemichannels produce aberrant channel activity, which gives rise to deafness and skin disorders, including keratitis-ichthyosis-deafness (KID) syndrome. Structural and functional studies indicate that the NT of connex...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Rockefeller University Press
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6400520/ https://www.ncbi.nlm.nih.gov/pubmed/30530766 http://dx.doi.org/10.1085/jgp.201812219 |
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author | Valdez Capuccino, Juan M. Chatterjee, Payal García, Isaac E. Botello-Smith, Wesley M. Zhang, Han Harris, Andrew L. Luo, Yun Contreras, Jorge E. |
author_facet | Valdez Capuccino, Juan M. Chatterjee, Payal García, Isaac E. Botello-Smith, Wesley M. Zhang, Han Harris, Andrew L. Luo, Yun Contreras, Jorge E. |
author_sort | Valdez Capuccino, Juan M. |
collection | PubMed |
description | A group of human mutations within the N-terminal (NT) domain of connexin 26 (Cx26) hemichannels produce aberrant channel activity, which gives rise to deafness and skin disorders, including keratitis-ichthyosis-deafness (KID) syndrome. Structural and functional studies indicate that the NT of connexin hemichannels is folded into the pore, where it plays important roles in permeability and gating. In this study, we explore the molecular basis by which N14K, an NT KID mutant, promotes gain of function. In macroscopic and single-channel recordings, we find that the N14K mutant favors the open conformation of hemichannels, shifts calcium and voltage sensitivity, and slows deactivation kinetics. Multiple copies of MD simulations of WT and N14K hemichannels, followed by the Kolmogorov–Smirnov significance test (KS test) of the distributions of interaction energies, reveal that the N14K mutation significantly disrupts pairwise interactions that occur in WT hemichannels between residue K15 of one subunit and residue E101 of the adjacent subunit (E101 being located at the transition between transmembrane segment 2 [TM2] and the cytoplasmic loop [CL]). Double mutant cycle analysis supports coupling between the NT and the TM2/CL transition in WT hemichannels, which is disrupted in N14K mutant hemichannels. KS tests of the α carbon correlation coefficients calculated over MD trajectories suggest that the effects of the N14K mutation are not confined to the K15–E101 pairs but extend to essentially all pairwise residue correlations between the NT and TM2/CL interface. Together, our data indicate that the N14K mutation increases hemichannel open probability by disrupting interactions between the NT and the TM2/CL region of the adjacent connexin subunit. This suggests that NT–TM2/CL interactions facilitate Cx26 hemichannel closure. |
format | Online Article Text |
id | pubmed-6400520 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-64005202019-09-04 The connexin26 human mutation N14K disrupts cytosolic intersubunit interactions and promotes channel opening Valdez Capuccino, Juan M. Chatterjee, Payal García, Isaac E. Botello-Smith, Wesley M. Zhang, Han Harris, Andrew L. Luo, Yun Contreras, Jorge E. J Gen Physiol Research Articles A group of human mutations within the N-terminal (NT) domain of connexin 26 (Cx26) hemichannels produce aberrant channel activity, which gives rise to deafness and skin disorders, including keratitis-ichthyosis-deafness (KID) syndrome. Structural and functional studies indicate that the NT of connexin hemichannels is folded into the pore, where it plays important roles in permeability and gating. In this study, we explore the molecular basis by which N14K, an NT KID mutant, promotes gain of function. In macroscopic and single-channel recordings, we find that the N14K mutant favors the open conformation of hemichannels, shifts calcium and voltage sensitivity, and slows deactivation kinetics. Multiple copies of MD simulations of WT and N14K hemichannels, followed by the Kolmogorov–Smirnov significance test (KS test) of the distributions of interaction energies, reveal that the N14K mutation significantly disrupts pairwise interactions that occur in WT hemichannels between residue K15 of one subunit and residue E101 of the adjacent subunit (E101 being located at the transition between transmembrane segment 2 [TM2] and the cytoplasmic loop [CL]). Double mutant cycle analysis supports coupling between the NT and the TM2/CL transition in WT hemichannels, which is disrupted in N14K mutant hemichannels. KS tests of the α carbon correlation coefficients calculated over MD trajectories suggest that the effects of the N14K mutation are not confined to the K15–E101 pairs but extend to essentially all pairwise residue correlations between the NT and TM2/CL interface. Together, our data indicate that the N14K mutation increases hemichannel open probability by disrupting interactions between the NT and the TM2/CL region of the adjacent connexin subunit. This suggests that NT–TM2/CL interactions facilitate Cx26 hemichannel closure. Rockefeller University Press 2019-03-04 /pmc/articles/PMC6400520/ /pubmed/30530766 http://dx.doi.org/10.1085/jgp.201812219 Text en © 2019 Valdez Capuccino et al. http://www.rupress.org/terms/https://creativecommons.org/licenses/by-nc-sa/4.0/This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Research Articles Valdez Capuccino, Juan M. Chatterjee, Payal García, Isaac E. Botello-Smith, Wesley M. Zhang, Han Harris, Andrew L. Luo, Yun Contreras, Jorge E. The connexin26 human mutation N14K disrupts cytosolic intersubunit interactions and promotes channel opening |
title | The connexin26 human mutation N14K disrupts cytosolic intersubunit interactions and promotes channel opening |
title_full | The connexin26 human mutation N14K disrupts cytosolic intersubunit interactions and promotes channel opening |
title_fullStr | The connexin26 human mutation N14K disrupts cytosolic intersubunit interactions and promotes channel opening |
title_full_unstemmed | The connexin26 human mutation N14K disrupts cytosolic intersubunit interactions and promotes channel opening |
title_short | The connexin26 human mutation N14K disrupts cytosolic intersubunit interactions and promotes channel opening |
title_sort | connexin26 human mutation n14k disrupts cytosolic intersubunit interactions and promotes channel opening |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6400520/ https://www.ncbi.nlm.nih.gov/pubmed/30530766 http://dx.doi.org/10.1085/jgp.201812219 |
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