Membrane topology of an ATP-gated ion channel (P2X receptor).

Détails

ID Serval
serval:BIB_5C2DB6D4A682
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Membrane topology of an ATP-gated ion channel (P2X receptor).
Périodique
Journal of Biological Chemistry
Auteur(s)
Newbolt A., Stoop R., Virginio C., Surprenant A., North R.A., Buell G., Rassendren F.
ISSN
0021-9258 (Print)
ISSN-L
0021-9258
Statut éditorial
Publié
Date de publication
1998
Peer-reviewed
Oui
Volume
273
Numéro
24
Pages
15177-15182
Langue
anglais
Notes
Publication types: Journal ArticlePublication Status: ppublish
Résumé
Western blots of Xenopus oocyte membrane preparations showed that the apparent molecular mass of the wild type P2X2 receptor (about 65 kDa) was reduced by pretreatment with endoglycosidase H. Mutagenesis of one or more of three potential asparagines (N182S, N239S, and N298S) followed by Western blots showed that each of the sites was glycosylated in the wild type receptor. Functional channels were formed by receptors lacking any single asparagine, but not by channels mutated in two or three positions. Artificial consensus sequences (N-X-S/T) introduced into the N-terminal region (asparagine at position 9, 16, or 26) were not glycosylated. Asparagines were glycosylated when introduced at the C-terminal end of the first hydrophobic domain (positions 62 and 66) and at the N-terminal end of the second hydrophobic domain (position 324). A protein in which the C terminus of one P2X2 subunit was joined to the N terminus of a second P2X2 subunit (from a concatenated cDNA) had twice the molecular mass of the P2X2 receptor subunit, and formed fully functional channels. The experiments provide direct evidence for the topology originally proposed for the P2X receptor, with intracellular N and C termini, two membrane-spanning domains, and a large extracellular loop.
Mots-clé
Adenosine Triphosphate/pharmacology, Amino Acid Sequence, Animals, Cell Line, Dimerization, Electrophysiology, Glycosylation, Hexosaminidases/metabolism, Humans, Ion Channel Gating/physiology, Membrane Proteins/chemistry, Molecular Sequence Data, Molecular Weight, Mutagenesis, Site-Directed/genetics, Oocytes/physiology, Receptors, Purinergic P2/chemistry, Receptors, Purinergic P2X2, Transfection/genetics, Xenopus laevis
Pubmed
Création de la notice
29/10/2013 11:14
Dernière modification de la notice
20/08/2019 14:14
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