2026-04-13T01:43:55Zhttps://recercat.cat/oai/request

oai:recercat.cat:2445/2285312026-04-10T21:57:45Zcom_2072_1057col_2072_478780col_2072_478799col_2072_478858col_2072_478916col_2072_478917

00925njm 22002777a 4500 dc Gaitán-Peñas, Héctor author Pérez González, Anna Priscil·la author González Subías, Marc author Zdebik, Anselm A. author Gasull Casanova, Xavier author Buey, Ruben M. author Errasti-Murugarren, Ekaitz author Estévez Povedano, Raúl author 2026-03-26T13:55:39Z 2026-03-26T13:55:39Z 2025-02-07 2026-03-26T13:55:41Z 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. Estructura molecular Conformació de proteïnes Biologia molecular Molecular structure Proteins conformation Molecular biology Identification of a crosstalk between ClC-1 C-terminal CBS domains and the transmembrane region