Cargando…
The Mechanism of Fast-Gate Opening in ClC-0
ClC-0 is a chloride channel whose gating is sensitive to both voltage and chloride. Based on analysis of gating kinetics using single-channel recordings, a five-state model was proposed to describe the dependence of ClC-0 fast-gate opening on voltage and external chloride (Chen, T.-Y., and C. Miller...
Autores principales: | , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2007
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2151655/ https://www.ncbi.nlm.nih.gov/pubmed/17846164 http://dx.doi.org/10.1085/jgp.200709759 |
_version_ | 1782144764513091584 |
---|---|
author | Engh, Anita M. Faraldo-Gómez, José D. Maduke, Merritt |
author_facet | Engh, Anita M. Faraldo-Gómez, José D. Maduke, Merritt |
author_sort | Engh, Anita M. |
collection | PubMed |
description | ClC-0 is a chloride channel whose gating is sensitive to both voltage and chloride. Based on analysis of gating kinetics using single-channel recordings, a five-state model was proposed to describe the dependence of ClC-0 fast-gate opening on voltage and external chloride (Chen, T.-Y., and C. Miller. 1996. J. Gen. Physiol. 108:237–250). We aimed to use this five-state model as a starting point for understanding the structural changes that occur during gating. Using macroscopic patch recordings, we were able to reproduce the effects of voltage and chloride that were reported by Chen and Miller and to fit our opening rate constant data to the five-state model. Upon further analysis of both our data and those of Chen and Miller, we learned that in contrast to their conclusions, (a) the features in the data are not adequate to rule out a simpler four-state model, and (b) the chloride-binding step is voltage dependent. In order to be able to evaluate the effects of mutants on gating (described in the companion paper, see Engh et al. on p. 351 of this issue), we developed a method for determining the error on gating model parameters, and evaluated the sources of this error. To begin to mesh the kinetic model(s) with the known CLC structures, a model of ClC-0 was generated computationally based on the X-ray crystal structure of the prokaryotic homolog ClC-ec1. Analysis of pore electrostatics in this homology model suggests that at least two of the conclusions derived from the gating kinetics analysis are consistent with the known CLC structures: (1) chloride binding is necessary for channel opening, and (2) chloride binding to any of the three known chloride-binding sites must be voltage dependent. |
format | Text |
id | pubmed-2151655 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21516552008-04-01 The Mechanism of Fast-Gate Opening in ClC-0 Engh, Anita M. Faraldo-Gómez, José D. Maduke, Merritt J Gen Physiol Articles ClC-0 is a chloride channel whose gating is sensitive to both voltage and chloride. Based on analysis of gating kinetics using single-channel recordings, a five-state model was proposed to describe the dependence of ClC-0 fast-gate opening on voltage and external chloride (Chen, T.-Y., and C. Miller. 1996. J. Gen. Physiol. 108:237–250). We aimed to use this five-state model as a starting point for understanding the structural changes that occur during gating. Using macroscopic patch recordings, we were able to reproduce the effects of voltage and chloride that were reported by Chen and Miller and to fit our opening rate constant data to the five-state model. Upon further analysis of both our data and those of Chen and Miller, we learned that in contrast to their conclusions, (a) the features in the data are not adequate to rule out a simpler four-state model, and (b) the chloride-binding step is voltage dependent. In order to be able to evaluate the effects of mutants on gating (described in the companion paper, see Engh et al. on p. 351 of this issue), we developed a method for determining the error on gating model parameters, and evaluated the sources of this error. To begin to mesh the kinetic model(s) with the known CLC structures, a model of ClC-0 was generated computationally based on the X-ray crystal structure of the prokaryotic homolog ClC-ec1. Analysis of pore electrostatics in this homology model suggests that at least two of the conclusions derived from the gating kinetics analysis are consistent with the known CLC structures: (1) chloride binding is necessary for channel opening, and (2) chloride binding to any of the three known chloride-binding sites must be voltage dependent. The Rockefeller University Press 2007-10 /pmc/articles/PMC2151655/ /pubmed/17846164 http://dx.doi.org/10.1085/jgp.200709759 Text en Copyright © 2007, The Rockefeller University Press 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 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Articles Engh, Anita M. Faraldo-Gómez, José D. Maduke, Merritt The Mechanism of Fast-Gate Opening in ClC-0 |
title | The Mechanism of Fast-Gate Opening in ClC-0 |
title_full | The Mechanism of Fast-Gate Opening in ClC-0 |
title_fullStr | The Mechanism of Fast-Gate Opening in ClC-0 |
title_full_unstemmed | The Mechanism of Fast-Gate Opening in ClC-0 |
title_short | The Mechanism of Fast-Gate Opening in ClC-0 |
title_sort | mechanism of fast-gate opening in clc-0 |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2151655/ https://www.ncbi.nlm.nih.gov/pubmed/17846164 http://dx.doi.org/10.1085/jgp.200709759 |
work_keys_str_mv | AT enghanitam themechanismoffastgateopeninginclc0 AT faraldogomezjosed themechanismoffastgateopeninginclc0 AT madukemerritt themechanismoffastgateopeninginclc0 AT enghanitam mechanismoffastgateopeninginclc0 AT faraldogomezjosed mechanismoffastgateopeninginclc0 AT madukemerritt mechanismoffastgateopeninginclc0 |