دورية أكاديمية
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Mapping the molecular motions of 5-HT 3 serotonin-gated channel by voltage-clamp fluorometry.

التفاصيل البيبلوغرافية
العنوان: Mapping the molecular motions of 5-HT 3 serotonin-gated channel by voltage-clamp fluorometry.
المؤلفون: Peverini L; Institut Pasteur, Université Paris Cité, CNRS UMR 3571, Channel-Receptors Unit, Paris, France., Shi S; Institut Pasteur, Université Paris Cité, CNRS UMR 3571, Channel-Receptors Unit, Paris, France., Medjebeur K; Institut Pasteur, Université Paris Cité, CNRS UMR 3571, Channel-Receptors Unit, Paris, France., Corringer PJ; Institut Pasteur, Université Paris Cité, CNRS UMR 3571, Channel-Receptors Unit, Paris, France.
المصدر: ELife [Elife] 2024 Jun 24; Vol. 12. Date of Electronic Publication: 2024 Jun 24.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: eLife Sciences Publications, Ltd Country of Publication: England NLM ID: 101579614 Publication Model: Electronic Cited Medium: Internet ISSN: 2050-084X (Electronic) Linking ISSN: 2050084X NLM ISO Abbreviation: Elife Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Cambridge, UK : eLife Sciences Publications, Ltd., 2012-
مواضيع طبية MeSH: Receptors, Serotonin, 5-HT3*/metabolism , Receptors, Serotonin, 5-HT3*/chemistry , Receptors, Serotonin, 5-HT3*/genetics , Fluorometry*/methods , Patch-Clamp Techniques* , Protein Conformation*, Humans ; Serotonin/metabolism ; Cryoelectron Microscopy ; HEK293 Cells ; Binding Sites ; Ion Channel Gating
مستخلص: The serotonin-gated ion channel (5-HT 3 R) mediates excitatory neuronal communication in the gut and the brain. It is the target for setrons, a class of competitive antagonists widely used as antiemetics, and is involved in several neurological diseases. Cryo-electron microscopy (cryo-EM) of the 5-HT 3 R in complex with serotonin or setrons revealed that the protein has access to a wide conformational landscape. However, assigning known high-resolution structures to actual states contributing to the physiological response remains a challenge. In the present study, we used voltage-clamp fluorometry (VCF) to measure simultaneously, for 5-HT 3 R expressed at a cell membrane, conformational changes by fluorescence and channel opening by electrophysiology. Four positions identified by mutational screening report motions around and outside the serotonin-binding site through incorporation of cysteine-tethered rhodamine dyes with or without a nearby quenching tryptophan. VCF recordings show that the 5-HT 3 R has access to four families of conformations endowed with distinct fluorescence signatures: 'resting-like' without ligand, 'inhibited-like' with setrons, 'pre-active-like' with partial agonists, and 'active-like' (open channel) with partial and strong agonists. Data are remarkably consistent with cryo-EM structures, the fluorescence partners matching respectively apo, setron-bound, 5-HT bound-closed, and 5-HT-bound-open conformations. Data show that strong agonists promote a concerted motion of all fluorescently labeled sensors during activation, while partial agonists, especially when loss-of-function mutations are engineered, stabilize both active and pre-active conformations. In conclusion, VCF, though the monitoring of electrophysiologically silent conformational changes, illuminates allosteric mechanisms contributing to signal transduction and their differential regulation by important classes of physiological and clinical effectors.
Competing Interests: LP, SS, KM, PC No competing interests declared
(© 2023, Peverini et al.)
References: J Biol Chem. 2009 Jun 5;284(23):15847-56. (PMID: 19286654)
Nat Rev Gastroenterol Hepatol. 2013 Aug;10(8):473-86. (PMID: 23797870)
Nature. 2000 Feb 17;403(6771):773-6. (PMID: 10693806)
J Neurosci. 2004 Dec 1;24(48):10924-40. (PMID: 15574743)
Science. 1996 Jan 12;271(5246):213-6. (PMID: 8539623)
Nat Commun. 2019 Jul 19;10(1):3225. (PMID: 31324772)
J Neurochem. 2008 Apr;105(2):413-24. (PMID: 18047559)
Subcell Biochem. 2021;96:373-408. (PMID: 33252737)
Elife. 2020 Oct 16;9:. (PMID: 33063666)
Nat Commun. 2023 Feb 13;14(1):795. (PMID: 36781912)
Nature. 2018 Nov;563(7730):270-274. (PMID: 30401837)
J Neurosci. 2012 Jan 25;32(4):1336-52. (PMID: 22279218)
Cell. 2021 Feb 18;184(4):957-968.e21. (PMID: 33567265)
J Biol Chem. 2007 Dec 7;282(49):36057-67. (PMID: 17911099)
Nature. 2009 May 21;459(7245):451-4. (PMID: 19339970)
Eur J Pharmacol. 1990 Jun 21;182(1):193-7. (PMID: 2144822)
Nature. 2014 Aug 21;512(7514):276-81. (PMID: 25119048)
Nat Commun. 2018 Feb 6;9(1):514. (PMID: 29410406)
Nat Commun. 2021 Feb 16;12(1):1074. (PMID: 33594077)
Biochemistry. 2008 Jun 17;47(24):6370-7. (PMID: 18498149)
Biophys J. 2009 Sep 2;97(5):1335-45. (PMID: 19720021)
J Mol Biol. 2021 Aug 20;433(17):167128. (PMID: 34224751)
J Physiol. 2007 May 15;581(Pt 1):51-73. (PMID: 17331994)
ACS Chem Neurosci. 2013 Feb 20;4(2):248-54. (PMID: 23421675)
Neuropsychopharmacology. 2022 Mar;47(4):880-890. (PMID: 35017671)
Mol Pharmacol. 2019 Dec;96(6):720-734. (PMID: 31582575)
Nature. 2018 Nov;563(7730):275-279. (PMID: 30401839)
Elife. 2017 Mar 15;6:. (PMID: 28294942)
Nat Commun. 2020 Oct 23;11(1):5369. (PMID: 33097732)
Mol Membr Biol. 2006 Sep-Oct;23(5):442-51. (PMID: 17060161)
J Neurosci. 2004 Oct 13;24(41):9097-104. (PMID: 15483128)
Elife. 2021 Sep 30;10:. (PMID: 34590583)
J Neurochem. 2009 Mar;108(6):1585-94. (PMID: 19166513)
Neuron. 2016 May 4;90(3):452-70. (PMID: 27151638)
Proc Natl Acad Sci U S A. 2004 Jul 6;101(27):10195-200. (PMID: 15218096)
Nat Struct Mol Biol. 2024 Apr;31(4):598-609. (PMID: 38177669)
Structure. 2020 Oct 6;28(10):1131-1140.e4. (PMID: 32726573)
Pharmacol Rev. 2019 Jul;71(3):383-412. (PMID: 31243157)
J Pharmacol Exp Ther. 2011 Oct;339(1):125-31. (PMID: 21775477)
J Mol Biol. 1965 May;12:88-118. (PMID: 14343300)
Curr Pharm Des. 2006;12(28):3615-30. (PMID: 17073663)
Biochemistry. 2010 Nov 16;49(45):9722-31. (PMID: 20886836)
معلومات مُعتمدة: 788974 International ERC_ European Research Council
فهرسة مساهمة: Keywords: 5-HT3 receptors; allosteric mechanisms; biochemistry; chemical biology; mouse; neuropharmacology; voltage-clamp fluorometry
المشرفين على المادة: 0 (Receptors, Serotonin, 5-HT3)
333DO1RDJY (Serotonin)
تواريخ الأحداث: Date Created: 20240624 Date Completed: 20240624 Latest Revision: 20240711
رمز التحديث: 20240711
مُعرف محوري في PubMed: PMC11196107
DOI: 10.7554/eLife.93174
PMID: 38913422
قاعدة البيانات: MEDLINE
الوصف
تدمد:2050-084X
DOI:10.7554/eLife.93174