دورية أكاديمية
أعرض في EDS

Thermal unfolding of a mammalian pentameric ligand-gated ion channel proceeds at consecutive, distinct steps.

التفاصيل البيبلوغرافية
العنوان: Thermal unfolding of a mammalian pentameric ligand-gated ion channel proceeds at consecutive, distinct steps.
المؤلفون: Tol MB; Laboratory of Physical Chemistry of Polymers and Membranes, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland., Deluz C, Hassaine G, Graff A, Stahlberg H, Vogel H
المصدر: The Journal of biological chemistry [J Biol Chem] 2013 Feb 22; Vol. 288 (8), pp. 5756-69. Date of Electronic Publication: 2012 Dec 29.
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE
أسماء مطبوعة: Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology
مواضيع طبية MeSH: Ligand-Gated Ion Channels/*metabolism , Receptors, Serotonin, 5-HT2/*metabolism, Animals ; CHO Cells ; Cell Membrane/metabolism ; Cricetinae ; DNA, Complementary/metabolism ; Detergents/chemistry ; Detergents/pharmacology ; Hot Temperature ; Ligands ; Lipid Bilayers/chemistry ; Mice ; Microscopy, Electron, Transmission/methods ; Microscopy, Fluorescence/methods ; Models, Biological ; Protein Denaturation ; Protein Structure, Tertiary ; Spectrometry, Fluorescence/methods ; Temperature
مستخلص: Pentameric ligand-gated ion channels (LGICs) play an important role in fast synaptic signal transduction. Binding of agonists to the β-sheet-structured extracellular domain opens an ion channel in the transmembrane α-helical region of the LGIC. How the structurally distinct and distant domains are functionally coupled for such central transmembrane signaling processes remains an open question. To obtain detailed information about the stability of and the coupling between these different functional domains, we analyzed the thermal unfolding of a homopentameric LGIC, the 5-hydroxytryptamine receptor (ligand binding, secondary structure, accessibility of Trp and Cys residues, and aggregation), in plasma membranes as well as during detergent extraction, purification, and reconstitution into artificial lipid bilayers. We found a large loss in thermostability correlating with the loss of the lipid bilayer during membrane solubilization and purification. Thermal unfolding of the 5-hydroxytryptamine receptor occurred in consecutive steps at distinct protein locations. A loss of ligand binding was detected first, followed by formation of different transient low oligomeric states of receptor pentamers, followed by partial unfolding of helical parts of the protein, which finally lead to the formation receptor aggregates. Structural destabilization of the receptor in detergents could be partially reversed by reconstituting the receptor into lipid bilayers. Our results are important because they quantify the stability of LGICs during detergent extraction and purification and can be used to create stabilized receptor proteins for structural and functional studies.
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المشرفين على المادة: 0 (DNA, Complementary)
0 (Detergents)
0 (Ligand-Gated Ion Channels)
0 (Ligands)
0 (Lipid Bilayers)
0 (Receptors, Serotonin, 5-HT2)
تواريخ الأحداث: Date Created: 20130101 Date Completed: 20130425 Latest Revision: 20211021
رمز التحديث: 20240628
مُعرف محوري في PubMed: PMC3581420
DOI: 10.1074/jbc.M112.422287
PMID: 23275379
قاعدة البيانات: MEDLINE
الوصف
تدمد:1083-351X
DOI:10.1074/jbc.M112.422287