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Similar molecular determinants on Rem mediate two distinct modes of inhibition of Ca(V)1.2 channels

Rad/Rem/Rem2/Gem (RGK) proteins are Ras-like GTPases that potently inhibit all high-voltage-gated calcium (Ca(V)1/Ca(V)2) channels and are, thus, well-positioned to tune diverse physiological processes. Understanding how RGK proteins inhibit Ca(V) channels is important for perspectives on their (pat...

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Autores principales: Puckerin, Akil A., Chang, Donald D., Subramanyam, Prakash, Colecraft, Henry M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4988437/
https://www.ncbi.nlm.nih.gov/pubmed/27115600
http://dx.doi.org/10.1080/19336950.2016.1180489
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author Puckerin, Akil A.
Chang, Donald D.
Subramanyam, Prakash
Colecraft, Henry M.
author_facet Puckerin, Akil A.
Chang, Donald D.
Subramanyam, Prakash
Colecraft, Henry M.
author_sort Puckerin, Akil A.
collection PubMed
description Rad/Rem/Rem2/Gem (RGK) proteins are Ras-like GTPases that potently inhibit all high-voltage-gated calcium (Ca(V)1/Ca(V)2) channels and are, thus, well-positioned to tune diverse physiological processes. Understanding how RGK proteins inhibit Ca(V) channels is important for perspectives on their (patho)physiological roles and could advance their development and use as genetically-encoded Ca(V) channel blockers. We previously reported that Rem can block surface Ca(V)1.2 channels in 2 independent ways that engage distinct components of the channel complex: (1) by binding auxiliary β subunits (β-binding-dependent inhibition, or BBD); and (2) by binding the pore-forming α(1C) subunit N-terminus (α(1C)-binding-dependent inhibition, or ABD). By contrast, Gem uses only the BBD mechanism to block Ca(V)1.2. Rem molecular determinants required for BBD Ca(V)1.2 inhibition are the distal C-terminus and the guanine nucleotide binding G-domain which interact with the plasma membrane and Ca(V)β, respectively. However, Rem determinants for ABD Ca(V)1.2 inhibition are unknown. Here, combining fluorescence resonance energy transfer, electrophysiology, systematic truncations, and Rem/Gem chimeras we found that the same Rem distal C-terminus and G-domain also mediate ABD Ca(V)1.2 inhibition, but with different interaction partners. Rem distal C-terminus interacts with α(1C) N-terminus to anchor the G-domain which likely interacts with an as-yet-unidentified site. In contrast to some previous studies, neither the C-terminus of Rem nor Gem was sufficient to inhibit Ca(V)1/Ca(V)2 channels. The results reveal that similar molecular determinants on Rem are repurposed to initiate 2 independent mechanisms of Ca(V)1.2 inhibition.
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spelling pubmed-49884372016-08-29 Similar molecular determinants on Rem mediate two distinct modes of inhibition of Ca(V)1.2 channels Puckerin, Akil A. Chang, Donald D. Subramanyam, Prakash Colecraft, Henry M. Channels (Austin) Research Paper Rad/Rem/Rem2/Gem (RGK) proteins are Ras-like GTPases that potently inhibit all high-voltage-gated calcium (Ca(V)1/Ca(V)2) channels and are, thus, well-positioned to tune diverse physiological processes. Understanding how RGK proteins inhibit Ca(V) channels is important for perspectives on their (patho)physiological roles and could advance their development and use as genetically-encoded Ca(V) channel blockers. We previously reported that Rem can block surface Ca(V)1.2 channels in 2 independent ways that engage distinct components of the channel complex: (1) by binding auxiliary β subunits (β-binding-dependent inhibition, or BBD); and (2) by binding the pore-forming α(1C) subunit N-terminus (α(1C)-binding-dependent inhibition, or ABD). By contrast, Gem uses only the BBD mechanism to block Ca(V)1.2. Rem molecular determinants required for BBD Ca(V)1.2 inhibition are the distal C-terminus and the guanine nucleotide binding G-domain which interact with the plasma membrane and Ca(V)β, respectively. However, Rem determinants for ABD Ca(V)1.2 inhibition are unknown. Here, combining fluorescence resonance energy transfer, electrophysiology, systematic truncations, and Rem/Gem chimeras we found that the same Rem distal C-terminus and G-domain also mediate ABD Ca(V)1.2 inhibition, but with different interaction partners. Rem distal C-terminus interacts with α(1C) N-terminus to anchor the G-domain which likely interacts with an as-yet-unidentified site. In contrast to some previous studies, neither the C-terminus of Rem nor Gem was sufficient to inhibit Ca(V)1/Ca(V)2 channels. The results reveal that similar molecular determinants on Rem are repurposed to initiate 2 independent mechanisms of Ca(V)1.2 inhibition. Taylor & Francis 2016-04-26 /pmc/articles/PMC4988437/ /pubmed/27115600 http://dx.doi.org/10.1080/19336950.2016.1180489 Text en © 2016 The Author(s). Published with license by Taylor & Francis Group, LLC http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License http://creativecommons.org/licenses/by-nc/3.0/, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.
spellingShingle Research Paper
Puckerin, Akil A.
Chang, Donald D.
Subramanyam, Prakash
Colecraft, Henry M.
Similar molecular determinants on Rem mediate two distinct modes of inhibition of Ca(V)1.2 channels
title Similar molecular determinants on Rem mediate two distinct modes of inhibition of Ca(V)1.2 channels
title_full Similar molecular determinants on Rem mediate two distinct modes of inhibition of Ca(V)1.2 channels
title_fullStr Similar molecular determinants on Rem mediate two distinct modes of inhibition of Ca(V)1.2 channels
title_full_unstemmed Similar molecular determinants on Rem mediate two distinct modes of inhibition of Ca(V)1.2 channels
title_short Similar molecular determinants on Rem mediate two distinct modes of inhibition of Ca(V)1.2 channels
title_sort similar molecular determinants on rem mediate two distinct modes of inhibition of ca(v)1.2 channels
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4988437/
https://www.ncbi.nlm.nih.gov/pubmed/27115600
http://dx.doi.org/10.1080/19336950.2016.1180489
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