The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface.

Détails

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Etat: Public
Version: de l'auteur
ID Serval
serval:BIB_76090C0DECB0
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
The Human Acid-Sensing Ion Channel ASIC1a: Evidence for a Homotetrameric Assembly State at the Cell Surface.
Périodique
Plos One
Auteur(s)
van Bemmelen M.X., Huser D., Gautschi I., Schild L.
ISSN
1932-6203 (Electronic)
ISSN-L
1932-6203
Statut éditorial
Publié
Date de publication
2015
Peer-reviewed
Oui
Volume
10
Numéro
8
Pages
e0135191
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: epublish
Résumé
The chicken acid-sensing ion channel ASIC1 has been crystallized as a homotrimer. We address here the oligomeric state of the functional ASIC1 in situ at the cell surface. The oligomeric states of functional ASIC1a and mutants with additional cysteines introduced in the extracellular pore vestibule were resolved on SDS-PAGE. The functional ASIC1 complexes were stabilized at the cell surface of Xenopus laevis oocytes or CHO cells either using the sulfhydryl crosslinker BMOE, or sodium tetrathionate (NaTT). Under these different crosslinking conditions ASIC1a migrates as four distinct oligomeric states that correspond by mass to multiples of a single ASIC1a subunit. The relative importance of each of the four ASIC1a oligomers was critically dependent on the availability of cysteines in the transmembrane domain for crosslinking, consistent with the presence of ASIC1a homo-oligomers. The expression of ASIC1a monomers, trimeric or tetrameric concatemeric cDNA constructs resulted in functional channels. The resulting ASIC1a complexes are resolved as a predominant tetramer over the other oligomeric forms, after stabilization with BMOE or NaTT and SDS-PAGE/western blot analysis. Our data identify a major ASIC1a homotetramer at the surface membrane of the cell expressing functional ASIC1a channel.
Mots-clé
Acid Sensing Ion Channels/chemistry, Animals, CHO Cells, Cell Membrane/chemistry, Cricetinae, Cricetulus, Cross-Linking Reagents/chemistry, Cysteine/chemistry, DNA, Complementary/chemistry, Humans, Mutagenesis, Site-Directed, Mutation, Oocytes/cytology, Protein Multimerization, Protein Structure, Tertiary, Tetrathionic Acid/chemistry, Xenopus laevis
Pubmed
Web of science
Open Access
Oui
Création de la notice
28/08/2015 17:27
Dernière modification de la notice
20/08/2019 14:33
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