Fast functional mapping of ligand-gated ion channels.
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
02 10 2023
02 10 2023
Historique:
received:
06
04
2023
accepted:
11
09
2023
medline:
4
10
2023
pubmed:
3
10
2023
entrez:
2
10
2023
Statut:
epublish
Résumé
Ligand-gated ion channels are formed by three to five subunits that control the opening of the pore in a cooperative fashion. We developed a microfluidic chip-based technique for studying ion currents and fluorescence signals in either excised membrane patches or whole cells to measure activation and deactivation kinetics of the channels as well as ligand binding and unbinding when using confocal patch-clamp fluorometry. We show how this approach produces in a few seconds either unidirectional concentration-activation relationships at or near equilibrium and, moreover, respective time courses of activation and deactivation for a large number of freely designed steps of the ligand concentration. The short measuring period strongly minimizes the contribution of disturbing superimposing effects such as run-down phenomena and desensitization effects. To validate gating mechanisms, complex kinetic schemes are quantified without the requirement to have data at equilibrium. The new method has potential for functionally analyzing any ligand-gated ion channel and, beyond, also for other receptors.
Identifiants
pubmed: 37783870
doi: 10.1038/s42003-023-05340-w
pii: 10.1038/s42003-023-05340-w
pmc: PMC10545696
doi:
Substances chimiques
Ligand-Gated Ion Channels
0
Ligands
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1003Informations de copyright
© 2023. Springer Nature Limited.
Références
J Cell Biol. 1990 Aug;111(2):599-606
pubmed: 2380245
Nature. 1997 Apr 10;386(6625):612-5
pubmed: 9121585
Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15485-90
pubmed: 8986838
Proc Natl Acad Sci U S A. 2009 Jul 7;106(27):11388-93
pubmed: 19549872
Anal Chem. 2004 Sep 1;76(17):4968-76
pubmed: 15373430
Nat Chem Biol. 2011 Dec 18;8(2):162-9
pubmed: 22179066
Anal Chem. 2010 Jun 1;82(11):4529-36
pubmed: 20443547
PLoS One. 2012;7(8):e42275
pubmed: 22879927
Pflugers Arch. 1981 Aug;391(2):85-100
pubmed: 6270629
Anal Chem. 2002 Dec 15;74(24):6133-8
pubmed: 12510730
Nature. 1998 Dec 17;396(6712):679-82
pubmed: 9872316
J Gen Physiol. 2012 May;139(5):333-48
pubmed: 22547664
J Gen Physiol. 2007 Aug;130(2):183-201
pubmed: 17664346
Biophys J. 1999 Aug;77(2):682-90
pubmed: 10423417
Annu Rev Physiol. 2001;63:235-57
pubmed: 11181956
Commun Biol. 2023 Jan 27;6(1):104
pubmed: 36707695
Mol Pharmacol. 1999 May;55(5):883-93
pubmed: 10220567
Nat Commun. 2013;4:2866
pubmed: 24287615
J Cell Physiol. 2016 Aug;231(8):1656-70
pubmed: 26627116
Nat Commun. 2016 Jan 08;7:10178
pubmed: 26744192
Nature. 1994 Nov 24;372(6504):369-74
pubmed: 7969497
Sci Signal. 2012 Aug 14;5(237):ra59
pubmed: 22894836
Neuron. 2010 Jul 15;67(1):75-85
pubmed: 20624593
Nature. 2012 May 10;485(7397):207-12
pubmed: 22535247
Nat Protoc. 2013;8(7):1299-306
pubmed: 23744290
Sci Rep. 2020 Dec 10;10(1):21751
pubmed: 33303878
Sci Rep. 2016 Jul 11;6:29378
pubmed: 27405959
Physiol Rev. 2002 Jul;82(3):769-824
pubmed: 12087135
Biophys J. 2007 Jul 1;93(1):74-91
pubmed: 17416622
Nature. 2017 Feb 2;542(7639):60-65
pubmed: 28099415
Nature. 2007 Mar 22;446(7134):440-3
pubmed: 17322905
Biophys J. 2019 Jun 18;116(12):2411-2422
pubmed: 31130235
Annu Rev Neurosci. 1996;19:235-63
pubmed: 8833443
Neuron. 2012 Jun 7;74(5):845-57
pubmed: 22681689
BMC Biophys. 2013 Mar 14;6:3
pubmed: 23497467
Elife. 2022 May 04;11:
pubmed: 35506659
Proc Natl Acad Sci U S A. 2019 May 14;116(20):10150-10155
pubmed: 31023886
Nature. 2009 Jul 30;460(7255):592-8
pubmed: 19641588
J Neurosci. 2017 Oct 4;37(40):9686-9704
pubmed: 28877967
Methods Mol Biol. 2014;1183:23-41
pubmed: 25023300
Neurosci Lett. 1987 Jun 15;77(2):199-204
pubmed: 2439957
J Biol Chem. 1977 Feb 10;252(3):864-5
pubmed: 14137