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PMID: 14523092 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Subunit arrangement in P2X receptors.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 23 ·No. 26 ·2003-10-01 ·Pages 8903-10

Jiang LH, Kim M, Spelta V, Bo X, Surprenant A, North RA

Abstract

ATP-gated ionotropic receptors (P2X receptors) are distributed widely in the nervous system. For example, a hetero-oligomeric receptor containing both P2X2 and P2X3 subunits is involved in primary afferent sensation. Each subunit has two membrane-spanning domains. We have used disulfide bond formation between engineered cysteines to demonstrate close proximity between the outer ends of the first transmembrane domain of one subunit and the second transmembrane domain of another. After expression in HEK 293 cells of such modified P2X2 or P2X4 subunits, the disulfide bond formation is evident because an ATP-evoked channel opening requires previous reduction with dithiothreitol. In the hetero-oligomeric P2X2/3 receptor the coexpression of doubly substituted subunits with wild-type partners allows us to deduce that the hetero-oligomeric channel contains adjacent P2X3 subunits but does not contain adjacent P2X2 subunits. The results suggest a "head-to-tail" subunit arrangement in the quaternary structure of P2X receptors and show that a trimeric P2X2/3 receptor would have the composition P2X2(P2X3)2.

MeSH Terms
Adenosine Triphosphate/analogs & derivatives,metabolism,pharmacology Animals Cell Line Disulfides/chemistry Humans Ion Channel Gating/drug effects,physiology Kidney/cytology,metabolism Ligands Macromolecular Substances Patch-Clamp Techniques Protein Engineering Protein Subunits/chemistry,genetics,physiology Rats Receptors, Purinergic P2/chemistry,genetics,physiology Receptors, Purinergic P2X Receptors, Purinergic P2X2 Receptors, Purinergic P2X3 Transfection
Chemicals
Disulfides Ligands Macromolecular Substances P2RX2 protein, human P2RX3 protein, human Protein Subunits Receptors, Purinergic P2 Receptors, Purinergic P2X Receptors, Purinergic P2X2 Receptors, Purinergic P2X3 Adenosine Triphosphate alpha,beta-methyleneadenosine 5'-triphosphate
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Jiang Lin-Hua
Institute of Molecular Physiology, University of Sheffield, Sheffield S10 2TN, United Kingdom.
Kim Miran
Spelta Valeria
Bo Xuenong
Surprenant Annmarie
North R Alan
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Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
Published
2003-10-01
Pages
8903-10
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6740386
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source PMC Full Text View journal details
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