We describe production of the human A(2A) adenosine receptor (A(2A)AR), a class A G protein-coupled receptor (GPCR) for ^(19)F-NMR and single-molecule fluorescence (SMF) spectroscopy. We explain in detail steps shared between the two sample preparation strategies, including expression and isolation of A(2A)AR and assembly of A(2A)AR in lipid nanodiscs and procedures for incorporation of either ^(19)F-NMR or fluorescence probes. Protocols for SMF experiments include sample setup, data...
[NMR paper] Synergies of Single Molecule Fluorescence and NMR for the Study of Intrinsically Disordered Proteins
Synergies of Single Molecule Fluorescence and NMR for the Study of Intrinsically Disordered Proteins
Single molecule fluorescence and nuclear magnetic resonance spectroscopy (NMR) are two very powerful techniques for the analysis of intrinsically disordered proteins (IDPs). Both techniques have individually made major contributions to deciphering the complex properties of IDPs and their interactions, and it has become evident that they can provide very complementary views on the distance-dynamics relationships of IDP systems. We now review the first approaches using both NMR and single...
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01-22-2022 02:38 AM
[NMR paper] Exploring Lipid and Membrane Protein Dynamics Using Lipid-Bilayer Nanodiscs and Solution-State NMR Spectroscopy.
Exploring Lipid and Membrane Protein Dynamics Using Lipid-Bilayer Nanodiscs and Solution-State NMR Spectroscopy.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--production.springer.de-OnlineResources-Logos-springerlink.gif Related Articles Exploring Lipid and Membrane Protein Dynamics Using Lipid-Bilayer Nanodiscs and Solution-State NMR Spectroscopy.
Methods Mol Biol. 2020;2127:397-419
Authors: Bibow S
Abstract
The relationship of membrane protein function and the surrounding lipid bilayer goes far beyond...
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03-03-2020 11:18 PM
[NMR paper] Single-Molecule Force Spectroscopy Trajectories of a Single Protein and Its Polyproteins Are Equivalent: A Direct Experimental Validation Based on A Small Protein NuG2
Single-Molecule Force Spectroscopy Trajectories of a Single Protein and Its Polyproteins Are Equivalent: A Direct Experimental Validation Based on A Small Protein NuG2
Single-molecule force spectroscopy (SMFS) has become a powerful tool in investigating the mechanical unfolding/folding of proteins at the single-molecule level. Polyproteins made of tandem identical repeats have been widely used in atomic force microscopy (AFM)-based SMFS studies, where polyproteins not only serve as fingerprints to identify single-molecule stretching events, but may also improve statistics of data...
Interaction of Human Chloride Intracellular ChannelProtein 1 (CLIC1) with Lipid Bilayers: A Fluorescence Study
Interaction of Human Chloride Intracellular ChannelProtein 1 (CLIC1) with Lipid Bilayers: A Fluorescence Study
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.6b00080/20160628/images/medium/bi-2016-00080v_0005.gif
Biochemistry
DOI: 10.1021/acs.biochem.6b00080
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06-30-2016 02:26 AM
[NMR paper] Gradual Disordering of the Native State on a Slow Two-State Folding Protein Monitored by Single-Molecule Fluorescence Spectroscopy and NMR.
Gradual Disordering of the Native State on a Slow Two-State Folding Protein Monitored by Single-Molecule Fluorescence Spectroscopy and NMR.
Gradual Disordering of the Native State on a Slow Two-State Folding Protein Monitored by Single-Molecule Fluorescence Spectroscopy and NMR.
J Phys Chem B. 2013 Jun 24;
Authors: Campos LA, Sadqi M, Liu J, Wang X, English DS, Munoz V
Abstract
Theory predicts that folding free energy landscapes are intrinsically malleable, and as such are expected to respond to perturbations in topographically complex...
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06-27-2013 01:52 AM
[NMR paper] Solution NMR spectroscopic characterization of human VDAC-2 in detergent micelles and lipid bilayer nanodiscs.
Solution NMR spectroscopic characterization of human VDAC-2 in detergent micelles and lipid bilayer nanodiscs.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Solution NMR spectroscopic characterization of human VDAC-2 in detergent micelles and lipid bilayer nanodiscs.
Biochim Biophys Acta. 2012 Jun;1818(6):1562-9
Authors: Yu TY, Raschle T, Hiller S, Wagner G
Abstract
Three isoforms of the human voltage-dependent anion channel (VDAC), located...
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04-04-2013 08:50 PM
Preparation of a functional GABARAP-lipid conjugate in nanodiscs and its investigation by solution NMR spectroscopy.
Preparation of a functional GABARAP-lipid conjugate in nanodiscs and its investigation by solution NMR spectroscopy.
Preparation of a functional GABARAP-lipid conjugate in nanodiscs and its investigation by solution NMR spectroscopy.
Chembiochem. 2010 Sep 24;11(14):1967-70
Authors: Ma P, Mohrlüder J, Schwarten M, Stoldt M, Singh SK, Hartmann R, Pacheco V, Willbold D
Production of human A2AAR in lipid nanodiscs for 19F-NMR and single-molecule fluorescence spectroscopy
Linear Mode
