[NMR paper] Label-free quantitative 1H NMR spectroscopy to study low-affinity ligand-protein interactions in solution: A contribution to the mechanism of polyphenol-mediated astringency.
Label-free quantitative 1H NMR spectroscopy to study low-affinity ligand-protein interactions in solution: A contribution to the mechanism of polyphenol-mediated astringency.
Label-free quantitative 1H NMR spectroscopy to study low-affinity ligand-protein interactions in solution: A contribution to the mechanism of polyphenol-mediated astringency.
PLoS One. 2017;12(9):e0184487
Authors: Delius J, Frank O, Hofmann T
Abstract
Nuclear magnetic resonance (NMR) spectroscopy is well-established in assessing the binding affinity between low molecular weight ligands and proteins. However, conventional NMR-based binding assays are often limited to small proteins of high purity and may require elaborate isotopic labeling of one of the potential binding partners. As protein-polyphenol complexation is assumed to be a key event in polyphenol-mediated oral astringency, here we introduce a label-free, ligand-focused 1H NMR titration assay to estimate binding affinities and characterize soluble complex formation between proteins and low molecular weight polyphenols. The method makes use of the effects of NMR line broadening due to protein-ligand interactions and quantitation of the non-bound ligand at varying protein concentrations by quantitative 1H NMR spectroscopy (qHNMR) using electronic reference to access in vivo concentration (ERETIC 2). This technique is applied to assess the interaction kinetics of selected astringent tasting polyphenols and purified mucin, a major lubricating glycoprotein of human saliva, as well as human whole saliva. The protein affinity values (BC50) obtained are subsequently correlated with the intrinsic mouth-puckering, astringent oral sensation imparted by these compounds. The quantitative NMR method is further exploited to study the effect of carboxymethyl cellulose, a candidate "anti-astringent" protein binding antagonist, on the polyphenol-protein interaction. Consequently, the NMR approach presented here proves to be a versatile tool to study the interactions between proteins and low-affinity ligands in solution and may find promising applications in the discovery of bioactives.
[NMR paper] Quantitative analysis of protein-ligand interactions by NMR.
Quantitative analysis of protein-ligand interactions by NMR.
Quantitative analysis of protein-ligand interactions by NMR.
Prog Nucl Magn Reson Spectrosc. 2016 Aug;96:47-57
Authors: Furukawa A, Konuma T, Yanaka S, Sugase K
Abstract
Protein-ligand interactions have been commonly studied through static structures of the protein-ligand complex. Recently, however, there has been increasing interest in investigating the dynamics of protein-ligand interactions both for fundamental understanding of the underlying mechanisms and for drug...
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08-31-2016 02:34 PM
Quantitative analysis of protein–ligand interactions by NMR
Quantitative analysis of protein–ligand interactions by NMR
Publication date: Available online 3 March 2016
Source:Progress in Nuclear Magnetic Resonance Spectroscopy</br>
Author(s): Ayako Furukawa, Tsuyoshi Konuma, Saeko Yanaka, Kenji Sugase</br>
Protein–ligand interactions have been commonly studied through static structures of the protein–ligand complex. Recently, however, there has been increasing interest in investigating the dynamics of protein–ligand interactions both for fundamental understanding of the underlying mechanisms and for drug development....
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03-03-2016 08:32 PM
Hyperpolarized Water to Study Protein–Ligand Interactions
From The DNP-NMR Blog:
Hyperpolarized Water to Study Protein–Ligand Interactions
Chappuis, Q., et al., Hyperpolarized Water to Study Protein–Ligand Interactions. The Journal of Physical Chemistry Letters, 2015. 6(9): p. 1674-1678.
http://dx.doi.org/10.1021/acs.jpclett.5b00403
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06-06-2015 12:01 PM
[NMR paper] The study of transient protein-nanoparticle interactions by solution NMR spectroscopy.
The study of transient protein-nanoparticle interactions by solution NMR spectroscopy.
Related Articles The study of transient protein-nanoparticle interactions by solution NMR spectroscopy.
Biochim Biophys Acta. 2015 Apr 30;
Authors: Assfalg M, Ragona L, Pagano K, D'Onofrio M, Zanzoni S, Tomaselli S, Molinari H
Abstract
The rapid development of novel nanoscale materials for applications in biomedicine urges an improved characterization of the nano-bio interfaces. Nanoparticles exhibit unique structures and properties, often...
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05-06-2015 11:59 AM
The study of transient protein-nanoparticle interactions by solution NMR spectroscopy
The study of transient protein-nanoparticle interactions by solution NMR spectroscopy
Publication date: Available online 30 April 2015
Source:Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics</br>
Author(s): Michael Assfalg , Laura Ragona , Katiuscia Pagano , Mariapina D’Onofrio , Serena Zanzoni , Simona Tomaselli , Henriette Molinari</br>
The rapid development of novel nanoscale materials for applications in biomedicine urges an improved characterization of the nano-bio interfaces. Nanoparticles exhibit unique structures and properties, often...
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04-30-2015 09:13 PM
[NMR paper] Analyzing protein-ligand interactions by dynamic NMR spectroscopy.
Analyzing protein-ligand interactions by dynamic NMR spectroscopy.
Related Articles Analyzing protein-ligand interactions by dynamic NMR spectroscopy.
Methods Mol Biol. 2013;1008:243-66
Authors: Mittermaier A, Meneses E
Abstract
Nuclear magnetic resonance (NMR) spectroscopy can provide detailed information on protein-ligand interactions that is inaccessible using other biophysical techniques. This chapter focuses on NMR-based approaches for extracting affinity and rate constants for weakly binding transient protein complexes with lifetimes...
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06-05-2013 06:53 PM
[NMR paper] NMR study of a membrane protein in detergent-free aqueous solution.
NMR study of a membrane protein in detergent-free aqueous solution.
Related Articles NMR study of a membrane protein in detergent-free aqueous solution.
Proc Natl Acad Sci U S A. 2005 Jun 21;102(25):8893-8
Authors: Zoonens M, Catoire LJ, Giusti F, Popot JL
One of the major obstacles to membrane protein (MP) structural studies is the destabilizing effect of detergents. Amphipols (APols) are short amphipathic polymers that can substitute for detergents to keep MPs water-soluble under mild conditions. In the present work, we have explored the...
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11-25-2010 08:21 PM
[NMR paper] Solution NMR study of DNA recognition mechanism of IRF4 protein.
Solution NMR study of DNA recognition mechanism of IRF4 protein.
Related Articles Solution NMR study of DNA recognition mechanism of IRF4 protein.
Nucleic Acids Symp Ser (Oxf). 2004;(48):105-6
Authors: Ishizaki I, Nomura M, Yamamoto K, Matsuyama T, Mishima M, Kojima C
Transcription factor IRF-4 prefers the DNA sequence including CCGAAA. The consensus sequence of the IRF family proteins is NNGAAA, and all crystal structures indicate the NN region does not interact with IRF proteins directly. Here the sequence preference of IRF-4 was...
Label-free quantitative 1H NMR spectroscopy to study low-affinity ligand-protein interactions in solution: A contribution to the mechanism of polyphenol-mediated astringency.
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