Determining the correct stoichiometry of Kv2.1/Kv6.4 heterotetramers, functional in multiple stoichiometrical configurations.
Animals
Antibodies
Cell Line
Fibroblasts
Gene Expression Regulation
HEK293 Cells
Humans
Membrane Potentials
Mice
Oocytes
/ metabolism
Photobleaching
Potassium
/ metabolism
Potassium Channels, Voltage-Gated
/ genetics
Receptor Protein-Tyrosine Kinases
/ genetics
Recombinant Proteins
Shab Potassium Channels
/ genetics
Xenopus
Kv channels
model selection
silent subunits
single subunit counting
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
28 04 2020
28 04 2020
Historique:
pubmed:
15
4
2020
medline:
11
8
2020
entrez:
15
4
2020
Statut:
ppublish
Résumé
The electrically silent (KvS) members of the voltage-gated potassium (Kv) subfamilies Kv5, Kv6, Kv8, and Kv9 selectively modulate Kv2 subunits by forming heterotetrameric Kv2/KvS channels. Based on the reported 3:1 stoichiometry of Kv2.1/Kv9.3 channels, we tested the hypothesis that Kv2.1/Kv6.4 channels express, in contrast to the assumed 3:1, in a 2:2 stoichiometry. We investigate the Kv2.1/Kv6.4 stoichiometry using single subunit counting and functional characterization of tetrameric concatemers. For selecting the most probable stoichiometry, we introduce a model-selection method that is applicable for any multimeric complex by investigating the stoichiometry of Kv2.1/Kv6.4 channels. Weighted likelihood calculations bring rigor to a powerful technique. Using the weighted-likelihood model-selection method and analysis of electrophysiological data, we show that Kv2.1/Kv6.4 channels express, in contrast to the assumed 3:1, in a 2:2 stoichiometry. Within this stoichiometry, the Kv6.4 subunits have to be positioned alternating with Kv2.1 to express functional channels. The variability in Kv2/KvS assembly increases the diversity of heterotetrameric configurations and extends the regulatory possibilities of KvS by allowing the presence of more than one silent subunit.
Identifiants
pubmed: 32284408
pii: 1916166117
doi: 10.1073/pnas.1916166117
pmc: PMC7196910
doi:
Substances chimiques
Antibodies
0
KCNB1 protein, human
0
KCNG4 protein, human
0
Kcnb1 protein, mouse
0
Potassium Channels, Voltage-Gated
0
Recombinant Proteins
0
Shab Potassium Channels
0
Ltk protein, mouse
EC 2.7.10.1
Receptor Protein-Tyrosine Kinases
EC 2.7.10.1
Potassium
RWP5GA015D
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
9365-9376Subventions
Organisme : CIHR
ID : MOP-136894
Pays : Canada
Commentaires et corrections
Type : CommentIn
Type : CommentIn
Déclaration de conflit d'intérêts
The authors declare no competing interest.
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