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Voltage-Sensing Domain of the Third Repeat of Human Skeletal Muscle NaV1.4 Channel As a New Target for Spider Gating Modifier Toxins
Voltage-gated sodium channels (NaV) have a modular architecture and contain five membrane domains. The central pore domain is responsible for ion conduction and contains a selectivity filter, while the four peripheral voltage-sensing domains (VSD-I/IV) are responsible for activation and rapid inacti...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
A.I. Gordeyev
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8084291/ https://www.ncbi.nlm.nih.gov/pubmed/33959393 http://dx.doi.org/10.32607/actanaturae.11279 |
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author | Myshkin, M. Yu. Paramonov, A. S. Kulbatskii, D. S. Surkova, E. A. Berkut, A. A. Vassilevski, A. A. Lyukmanova, E. N. Kirpichnikov, M. P. Shenkarev, Z. O. |
author_facet | Myshkin, M. Yu. Paramonov, A. S. Kulbatskii, D. S. Surkova, E. A. Berkut, A. A. Vassilevski, A. A. Lyukmanova, E. N. Kirpichnikov, M. P. Shenkarev, Z. O. |
author_sort | Myshkin, M. Yu. |
collection | PubMed |
description | Voltage-gated sodium channels (NaV) have a modular architecture and contain five membrane domains. The central pore domain is responsible for ion conduction and contains a selectivity filter, while the four peripheral voltage-sensing domains (VSD-I/IV) are responsible for activation and rapid inactivation of the channel. “Gating modifier” toxins from arthropod venoms interact with VSDs, influencing the activation and/or inactivation of the channel, and may serve as prototypes of new drugs for the treatment of various channelopathies and pain syndromes. The toxin-binding sites located on VSD-I, II and IV of mammalian NaV channels have been previously described. In this work, using the example of the Hm-3 toxin from the crab spider Heriaeus melloteei, we showed the presence of a toxin-binding site on VSD-III of the human skeletal muscle NaV1.4 channel. A developed cell-free protein synthesis system provided milligram quantities of isolated (separated from the channel) VSD-III and its 15N-labeled analogue. The interactions between VSD-III and Hm-3 were studied by NMR spectroscopy in the membrane-like environment of DPC/LDAO (1 : 1) micelles. Hm-3 has a relatively high affinity to VSD-III (dissociation constant of the complex Kd ~6 μM), comparable to the affinity to VSD‑I and exceeding the affinity to VSD-II. Within the complex, the positively charged Lys25 and Lys28 residues of the toxin probably interact with the S1–S2 extracellular loop of VSD-III. The Hm-3 molecule also contacts the lipid bilayer surrounding the channel. |
format | Online Article Text |
id | pubmed-8084291 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | A.I. Gordeyev |
record_format | MEDLINE/PubMed |
spelling | pubmed-80842912021-05-05 Voltage-Sensing Domain of the Third Repeat of Human Skeletal Muscle NaV1.4 Channel As a New Target for Spider Gating Modifier Toxins Myshkin, M. Yu. Paramonov, A. S. Kulbatskii, D. S. Surkova, E. A. Berkut, A. A. Vassilevski, A. A. Lyukmanova, E. N. Kirpichnikov, M. P. Shenkarev, Z. O. Acta Naturae Research Article Voltage-gated sodium channels (NaV) have a modular architecture and contain five membrane domains. The central pore domain is responsible for ion conduction and contains a selectivity filter, while the four peripheral voltage-sensing domains (VSD-I/IV) are responsible for activation and rapid inactivation of the channel. “Gating modifier” toxins from arthropod venoms interact with VSDs, influencing the activation and/or inactivation of the channel, and may serve as prototypes of new drugs for the treatment of various channelopathies and pain syndromes. The toxin-binding sites located on VSD-I, II and IV of mammalian NaV channels have been previously described. In this work, using the example of the Hm-3 toxin from the crab spider Heriaeus melloteei, we showed the presence of a toxin-binding site on VSD-III of the human skeletal muscle NaV1.4 channel. A developed cell-free protein synthesis system provided milligram quantities of isolated (separated from the channel) VSD-III and its 15N-labeled analogue. The interactions between VSD-III and Hm-3 were studied by NMR spectroscopy in the membrane-like environment of DPC/LDAO (1 : 1) micelles. Hm-3 has a relatively high affinity to VSD-III (dissociation constant of the complex Kd ~6 μM), comparable to the affinity to VSD‑I and exceeding the affinity to VSD-II. Within the complex, the positively charged Lys25 and Lys28 residues of the toxin probably interact with the S1–S2 extracellular loop of VSD-III. The Hm-3 molecule also contacts the lipid bilayer surrounding the channel. A.I. Gordeyev 2021 /pmc/articles/PMC8084291/ /pubmed/33959393 http://dx.doi.org/10.32607/actanaturae.11279 Text en Copyright ® 2021 National Research University Higher School of Economics. https://creativecommons.org/licenses/by/2.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Myshkin, M. Yu. Paramonov, A. S. Kulbatskii, D. S. Surkova, E. A. Berkut, A. A. Vassilevski, A. A. Lyukmanova, E. N. Kirpichnikov, M. P. Shenkarev, Z. O. Voltage-Sensing Domain of the Third Repeat of Human Skeletal Muscle NaV1.4 Channel As a New Target for Spider Gating Modifier Toxins |
title | Voltage-Sensing Domain of the Third Repeat of Human Skeletal Muscle NaV1.4 Channel As a New Target for Spider Gating Modifier Toxins |
title_full | Voltage-Sensing Domain of the Third Repeat of Human Skeletal Muscle NaV1.4 Channel As a New Target for Spider Gating Modifier Toxins |
title_fullStr | Voltage-Sensing Domain of the Third Repeat of Human Skeletal Muscle NaV1.4 Channel As a New Target for Spider Gating Modifier Toxins |
title_full_unstemmed | Voltage-Sensing Domain of the Third Repeat of Human Skeletal Muscle NaV1.4 Channel As a New Target for Spider Gating Modifier Toxins |
title_short | Voltage-Sensing Domain of the Third Repeat of Human Skeletal Muscle NaV1.4 Channel As a New Target for Spider Gating Modifier Toxins |
title_sort | voltage-sensing domain of the third repeat of human skeletal muscle nav1.4 channel as a new target for spider gating modifier toxins |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8084291/ https://www.ncbi.nlm.nih.gov/pubmed/33959393 http://dx.doi.org/10.32607/actanaturae.11279 |
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