Phospho-regulation, nucleotide binding and ion access control in potassium-chloride cotransporters.
HDX-MS
nucleotide binding
phospho-regulation
potassium-chloride co-transport
solute carrier
Journal
The EMBO journal
ISSN: 1460-2075
Titre abrégé: EMBO J
Pays: England
ID NLM: 8208664
Informations de publication
Date de publication:
15 07 2021
15 07 2021
Historique:
revised:
29
03
2021
received:
12
11
2020
accepted:
11
04
2021
pubmed:
26
5
2021
medline:
18
11
2021
entrez:
25
5
2021
Statut:
ppublish
Résumé
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, KCC1Δ19, 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.
Identifiants
pubmed: 34031912
doi: 10.15252/embj.2020107294
pmc: PMC8280820
doi:
Substances chimiques
Chlorides
0
Nucleotides
0
Symporters
0
Potassium
RWP5GA015D
Banques de données
PDB
['6Y5V', '7AIN', '7AIO', '7AIP', '7AIQ', '7AIR', '7NGB']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e107294Subventions
Organisme : Medical Research Council
ID : MC_PC_17137
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 060208/Z/00/Z
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_PC_17136
Pays : United Kingdom
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 093305/Z/10/Z
Pays : United Kingdom
Commentaires et corrections
Type : CommentIn
Informations de copyright
© 2021 The Authors. Published under the terms of the CC BY 4.0 license.
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