Related ArticlesProbing the Conformational States of Neurotensin Receptor 1 Variants*by NMR Site-Directed Methyl Labeling.
Chembiochem. 2020 Sep 02;:
Authors: Goba I, Goricanec D, Schum D, Hillenbrand M, Plückthun A, Hagn F
Abstract
G-protein coupled receptors are key players mediating signal transduction across the cell membrane. However, due to their intrinsic instability, many GPCRs are not suitable for structural investigations. Thus, various approaches have been developed in recent years to remedy this situation, including protein stabilization methods. This approach typically results in GPCRs that contain a varying number of mutations. Probing the functionality of such variants by*in vitro*and*in vivo*assays is often time-consuming and does not provide information on the conformational states that occur during activation. NMR spectroscopy has been proven to be a suitable technique to probe the conformational landscape of GPCRs in solution. Here, by using chemical modification of cysteine residues with an isotope-labeled methyl probe, we show that the conformational states of stabilized neurotensin receptor 1 variants can be efficiently assayed with 2D NMR experiments and correlated with their macroscopic activity profile. In addition, we show that the presented procedure can be utilized for the design of mutagenesis experiments to restore native-like conformational switching features in a stabilized receptor background. The presented chemical labeling approach will be broadly applicable in cases where low production yields do not permit uniform of amino-acid-selective labeling and where 1D NMR spectra of*19F-labeled receptors are not sufficient to resolve signal overlap.
PMID: 32881260 [PubMed - as supplied by publisher]
[NMR paper] Impact of polyphenols on receptor-ligand interactions by NMR: the case of neurotensin (NT)-neurotensin receptor fragment (NTS1) complex.
Impact of polyphenols on receptor-ligand interactions by NMR: the case of neurotensin (NT)-neurotensin receptor fragment (NTS1) complex.
Impact of polyphenols on receptor-ligand interactions by NMR: the case of neurotensin (NT)-neurotensin receptor fragment (NTS1) complex.
J Biomol Struct Dyn. 2019 May 03;:1-12
Authors: Richard T, Petit E, Pouységu L, Monti JP, Bondon A, Sylla T, Mérillon JM, Quideau S, Da Costa G
Abstract
Ligand-receptor interactions can be implicated in many pathological events such as chronic...
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[NMR paper] Optimization and 13CH3 methionine labeling of a signaling competent neurotensin receptor 1 variant for NMR studies.
Optimization and 13CH3 methionine labeling of a signaling competent neurotensin receptor 1 variant for NMR studies.
Optimization and 13CH3 methionine labeling of a signaling competent neurotensin receptor 1 variant for NMR studies.
Biochim Biophys Acta. 2018 Mar 26;:
Authors: Bumbak F, Keen AC, Gunn NJ, Gooley PR, Bathgate RAD, Scott DJ
Abstract
Neurotensin is a 13-residue peptide that acts as a neuromodulator of classical neurotransmitters such as dopamine and glutamate in the mammalian central nervous system, mainly by...
[NMR paper] Elucidation by NMR solution of neurotensin in small unilamellar vesicle environment: molecular surveys for neurotensin receptor recognition.
Elucidation by NMR solution of neurotensin in small unilamellar vesicle environment: molecular surveys for neurotensin receptor recognition.
Related Articles Elucidation by NMR solution of neurotensin in small unilamellar vesicle environment: molecular surveys for neurotensin receptor recognition.
J Biomol Struct Dyn. 2013;31(8):809-17
Authors: Da Costa G, Bondon A, Delalande O, Mouret L, Monti JP
Abstract
Neurotensin (NT) is a tridecapeptide, hormone in the periphery and neurotransmitter in the brain. We used high-resolution nuclear...
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02-08-2014 05:45 PM
Site-Directed Methyl Group Labeling as an NMR Probe of Structure and Dynamics in Supramolecular Protein Systems: Applications to the Proteasome and to the ClpP Protease
Site-Directed Methyl Group Labeling as an NMR Probe of Structure and Dynamics in Supramolecular Protein Systems: Applications to the Proteasome and to the ClpP Protease
Tomasz L. Religa, Amy M. Ruschak, Rina Rosenzweig and Lewis E. Kay
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja202259a/aop/images/medium/ja-2011-02259a_0002.gif
Journal of the American Chemical Society
DOI: 10.1021/ja202259a
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/rQfCMlQFoW8
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Site-Directed Methyl Group Labeling as an NMR Probe of Structure and Dynamics in Supra-Molecular Protein Systems: Applications to the Proteasome and to the ClpP Protease.
Site-Directed Methyl Group Labeling as an NMR Probe of Structure and Dynamics in Supra-Molecular Protein Systems: Applications to the Proteasome and to the ClpP Protease.
Site-Directed Methyl Group Labeling as an NMR Probe of Structure and Dynamics in Supra-Molecular Protein Systems: Applications to the Proteasome and to the ClpP Protease.
J Am Chem Soc. 2011 May 11;
Authors: Religa TL, Ruschak AM, Rosenzweig R, Kay LE
Methyl groups are powerful reporters of structure, motion and function in NMR studies of supra-molecular protein assemblies. Their...
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[NMR paper] Probing conformational disorder in neurotensin by two-dimensional solid-state NMR and comparison to molecular dynamics simulations.
Probing conformational disorder in neurotensin by two-dimensional solid-state NMR and comparison to molecular dynamics simulations.
Related Articles Probing conformational disorder in neurotensin by two-dimensional solid-state NMR and comparison to molecular dynamics simulations.
Biophys J. 2005 Sep;89(3):2113-20
Authors: Heise H, Luca S, de Groot BL, Grubmüller H, Baldus M
An approach is introduced to characterize conformational ensembles of intrinsically unstructured peptides on the atomic level using two-dimensional solid-state NMR data and...
Probing the Conformational States of Neurotensin Receptor 1 Variants*by NMR Site-Directed Methyl Labeling.
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