Phospho‐regulation, nucleotide binding and ion access control in potassium‐chloride cotransporters
| Autor: | Rebecca Ebenhoch, S.M.M. Mukhopadhyay, Samira Slimani, Henry Man, Gavin McKinley, Gabriella Reggiano, Patrizia Abrusci, Gamma Chi, Nicola A. Burgess-Brown, Kiran Bountra, Idlir Liko, Alexandre P. Garneau, Ben Tehan, Frank DiMaio, Katharina L. Dürr, Ali Jazayeri, Carol V. Robinson, Alejandra Fernández-Cid, Laurence E. Tremblay, Tina Bohstedt, Christophe P Moreau, Julie L. Lavoie, Xingyu Qiu, Haiping Tang, Dong Wang, Fernando G. Almeida, Paul Isenring |
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| Přispěvatelé: | Université de Montréal. Faculté de médecine. École de kinésiologie et des sciences de l'activité physique |
| Rok vydání: | 2021 |
| Předmět: |
Mutant
Biology General Biochemistry Genetics and Molecular Biology Cell Line 03 medical and health sciences 0302 clinical medicine Chlorides Adenine nucleotide Nucleotide binding Sf9 Cells Animals Humans News & Views Nucleotide Phosphorylation HDX-MS Molecular Biology Cell Size 030304 developmental biology chemistry.chemical_classification Potassium-chloride co-transport 0303 health sciences Symporters General Immunology and Microbiology Nucleotides General Neuroscience Solute carrier 3. Good health Solute carrier family Cell biology chemistry Cytoplasm Potassium Phospho-regulation Cotransporter 030217 neurology & neurosurgery Intracellular Signal Transduction |
| Zdroj: | The EMBO Journal 'EMBO Journal ', vol: 40, pages: e107294-1-e107294-20 (2021) EMBO J |
| ISSN: | 1460-2075 0261-4189 |
| DOI: | 10.15252/embj.2020107294 |
| Popis: | Potassium-coupled chloride transporters (KCCs) play crucial roles in regulating cell volume and intracellular chloride concentration. They are characteristically inhibited under isotonic conditions via phospho-regulatory sites located within the cytoplasmic termini. Decreased inhibitory phosphorylation in response to hypotonic cell swelling stimulates transport activity, and dysfunction of this regulatory process has been associated with various human diseases. Here, we present cryo-EM structures of human KCC3b and KCC1, revealing structural determinants for phospho-regulation in both N- and C-termini. We show that phospho-mimetic KCC3b is arrested in an inward-facing state in which intracellular ion access is blocked by extensive contacts with the N-terminus. In another mutant with increased isotonic transport activity, KCC1 Delta 19, this interdomain interaction is absent, likely due to a unique phospho-regulatory site in the KCC1 N-terminus. Furthermore, we map additional phosphorylation sites as well as a previously unknown ATP/ADP-binding pocket in the large C-terminal domain and show enhanced thermal stabilization of other CCCs by adenine nucleotides. These findings provide fundamentally new insights into the complex regulation of KCCs and may unlock innovative strategies for drug development. |
| Databáze: | OpenAIRE |
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