eprintid: 10205267
rev_number: 7
eprint_status: archive
userid: 699
dir: disk0/10/20/52/67
datestamp: 2025-02-26 08:33:52
lastmod: 2025-02-26 08:33:52
status_changed: 2025-02-26 08:33:52
type: article
metadata_visibility: show
sword_depositor: 699
creators_name: Gaitán-Peñas, Héctor
creators_name: Perez-Gonzalez, Anna Priscil·la
creators_name: González-Subías, Marc
creators_name: Zdebik, Anselm A
creators_name: Gasull, Xavier
creators_name: Buey, Rubén M
creators_name: Errasti-Murugarren, Ekaitz
creators_name: Estévez, Raúl
title: Identification of a crosstalk between ClC-1 C-terminal CBS domains and the transmembrane region
ispublished: inpress
divisions: UCL
divisions: B02
divisions: C08
divisions: D09
divisions: G02
keywords: ClC proteins; CBS domains; gating; intramolecular protein interaction
note: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
abstract: CLC channels and transporters have large C-terminal regions which contain two cystathionine β-synthetase (CBS) domains. It has been hypothesized that conformational changes in these domains upon nucleotide binding modulate the gating of the CLC dimer. It is not clear how rearrangements that occur in the CBS domains are transmitted to the ion pathway, as CBS domains interact with the rest of the channel at multiple locations and some of these sites are not visible in recent solved cryogenic electron microscopy structures or are difficult to model using the AlphaFold server. Using ClC-1 as a model, we started working with a described ClC-1 mutation (H835R) located in the first alpha helix of the CBS2 domain which changes the voltage dependence of gating. We then identified several residues located in the disorganized loop after helix R (R-linker) that revert the phenotype of this mutation. We additionally proved that R-linker's function is connected to the CBS2 domain as current intensity, plasma membrane levels and gating defects of several R-linker variants were corrected by adding the mutation H835R. Furthermore, cross-linking studies using newly developed split-cysless ClC-1 channels containing specific cysteine mutants in the R-linker and the CBS2 domain indicate that these two regions are in close contact. Considering these new results, we propose that conformational changes occurring in the CBS domains could be transmitted to the CLC intracellular chloride binding site by means of its interaction with the R-linker.
date: 2025-02-07
date_type: published
publisher: WILEY
official_url: https://doi.org/10.1113/jp287718
full_text_type: other
language: eng
verified: verified_manual
elements_id: 2362127
doi: 10.1113/JP287718
medium: Print-Electronic
lyricists_name: Zdebik, Anselm
lyricists_id: AAZDE01
actors_name: Zdebik, Anselm
actors_id: AAZDE01
actors_role: owner
funding_acknowledgements: [MICINN]; RTI2018-093493-B-I00; PID2021-126246NB-I00
full_text_status: restricted
publication: The Journal of Physiology
pages: 18
event_location: England
issn: 0022-3751
citation:        Gaitán-Peñas, Héctor;    Perez-Gonzalez, Anna Priscil·la;    González-Subías, Marc;    Zdebik, Anselm A;    Gasull, Xavier;    Buey, Rubén M;    Errasti-Murugarren, Ekaitz;           Gaitán-Peñas, Héctor;  Perez-Gonzalez, Anna Priscil·la;  González-Subías, Marc;  Zdebik, Anselm A;  Gasull, Xavier;  Buey, Rubén M;  Errasti-Murugarren, Ekaitz;  Estévez, Raúl;   - view fewer <#>    (2025)    Identification of a crosstalk between ClC-1 C-terminal CBS domains and the transmembrane region.                   The Journal of Physiology        10.1113/JP287718 <https://doi.org/10.1113/JP287718>.    (In press).   
 
document_url: https://discovery.ucl.ac.uk/id/eprint/10205267/1/18229_3_merged_1737545202.pdf