19F NMR as a Tool for Monitoring Individual Proteins in Complex Mixtures.
Mol Pharm. 2018 Jun 04;:
Authors: Edwards JM, Derrick JP, van der Walle CF, Golovanov AP
Abstract
The ability to monitor the behavior of individual proteins in complex mixtures has many potential uses, ranging from analysis of protein interactions in highly-concentrated solutions, modelling biological fluids or the intracellular environment, to optimizing biopharmaceutical co-formulations. Differential labelling NMR approaches, which tradi-tionally use 15N or 13C isotope incorporation during recombinant expression, are not always practical in cases when endogenous proteins are obtained from an organism, or where the expression system does not allow for efficient labelling, especially for larger proteins. This study proposes differential labelling of proteins by covalent attachment of 19F groups with distinct chemical shifts, giving each protein a unique spectral signature which can be monitored by 19F NMR without signal overlap, even in complex mixtures, and without any interfering signals from the buffer or other unlabelled components. Parameters such as signal intensities, translational diffusion coefficients and transverse relaxation rates, which report on the behaviour of individual proteins in the mixture, can be recorded even for proteins as large as antibodies at a wide range of concentrations.
PMID: 29863878 [PubMed - as supplied by publisher]
(3, 2)D 1 H, 13 C BIRD r,X -HSQC-TOCSY for NMR structure elucidation of mixtures: application to complex carbohydrates
(3, 2)D 1 H, 13 C BIRD r,X -HSQC-TOCSY for NMR structure elucidation of mixtures: application to complex carbohydrates
Abstract
Overlap of NMR signals is the major cause of difficulties associated with NMR structure elucidation of molecules contained in complex mixtures. A 2D homonuclear correlation spectroscopy in particular suffers from low dispersion of 1H chemical shifts; larger dispersion of 13C chemical shifts is often used to reduce this overlap, while still providing the protonā??proton correlation information e.g. in the form of a 2D 1H, 13C...
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01-11-2018 04:07 PM
[NMR paper] Fast 2D NMR Spectroscopy for In vivo Monitoring of Bacterial Metabolism in Complex Mixtures.
Fast 2D NMR Spectroscopy for In vivo Monitoring of Bacterial Metabolism in Complex Mixtures.
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Front Microbiol. 2017;8:1306
Authors: Dass R, Grudzia ? K, Ishikawa T, Nowakowski M, D?bowska R, Kazimierczuk K
Abstract
The biological toolbox is full of techniques developed originally for analytical chemistry. Among them, spectroscopic experiments are very important source of atomic-level structural information. Nuclear...
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08-05-2017 11:09 PM
[NMR paper] Expression, Purification, and Monitoring of Conformational Changes of hCB2 TMH67H8 in Different Membrane-Mimetic Lipid Mixtures Using Circular Dichroism and NMR Techniques.
Expression, Purification, and Monitoring of Conformational Changes of hCB2 TMH67H8 in Different Membrane-Mimetic Lipid Mixtures Using Circular Dichroism and NMR Techniques.
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Membranes (Basel). 2017 Feb 17;7(1):
Authors: Tiburu EK, Zhuang J, Fleischer HN, Arthur PK, Awandare GA
Abstract
This work was intended to develop self-assembly lipids for...
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02-22-2017 06:28 PM
[NMR paper] Monitoring the Interactions of a Ternary Complex Using NMR Spectroscopy: The Case of Sugars, Polyphenols, and Proteins.
Monitoring the Interactions of a Ternary Complex Using NMR Spectroscopy: The Case of Sugars, Polyphenols, and Proteins.
Monitoring the Interactions of a Ternary Complex Using NMR Spectroscopy: The Case of Sugars, Polyphenols, and Proteins.
Anal Chem. 2016 Dec 20;88(24):12470-12478
Authors: Faurie B, Dufourc EJ, Laguerre M, Pianet I
Abstract
Gaining insight into intermolecular interactions between multiple species is possible at an atomic level by looking at different parameters using different NMR techniques. In the specific...
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02-15-2017 03:40 PM
[NMR paper] NMRmix: A Tool for the Optimization of Compound Mixtures in 1D (1)H NMR Ligand Affinity Screens.
NMRmix: A Tool for the Optimization of Compound Mixtures in 1D (1)H NMR Ligand Affinity Screens.
Related Articles NMRmix: A Tool for the Optimization of Compound Mixtures in 1D (1)H NMR Ligand Affinity Screens.
J Proteome Res. 2016 Mar 11;
Authors: Stark JL, Eghbalnia HR, Lee W, Westler WM, Markley JL
Abstract
NMR ligand affinity screening is a powerful technique that is routinely used in drug discovery or functional genomics to directly detect protein-ligand binding events. Binding events can be identified by monitoring...
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03-12-2016 01:58 PM
Quantitative Trace Analysis of Complex Mixtures Using SABRE Hyperpolarization
From The DNP-NMR Blog:
Quantitative Trace Analysis of Complex Mixtures Using SABRE Hyperpolarization
Eshuis, N., et al., Quantitative Trace Analysis of Complex Mixtures Using SABRE Hyperpolarization. Angew Chem Int Ed Engl, 2014: p. n/a-n/a.
http://www.ncbi.nlm.nih.gov/pubmed/25469822
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01-12-2015 11:31 PM
Ultrafast quantitative 2D NMR: an efficient tool for the measurement of specific isotopic enrichments in complex biological mixtures.
Ultrafast quantitative 2D NMR: an efficient tool for the measurement of specific isotopic enrichments in complex biological mixtures.
Ultrafast quantitative 2D NMR: an efficient tool for the measurement of specific isotopic enrichments in complex biological mixtures.
Anal Chem. 2011 Apr 15;83(8):3112-9
Authors: Giraudeau P, Massou S, Robin Y, Cahoreau E, Portais JC, Akoka S
Two-dimensional nuclear magnetic resonance (2D NMR) is a promising tool for studying metabolic fluxes by measuring (13)C-enrichments in complex mixtures of (13)C-labeled...
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08-04-2011 11:41 AM
[NMR paper] NMR spectroscopic filtration of polypeptides and proteins in complex mixtures.
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J Biomol NMR. 2004 Aug;29(4):505-16
Authors: Rajagopalan S, Chow C, Raghunathan V, Fry CG, Cavagnero S
Due to the inherent complexity of the natural biological environment, most studies on polypeptides, proteins and nucleic acids have so far been performed in vitro, away from physiologically relevant conditions. Nuclear magnetic resonance is an ideal technique to extend...
19F NMR as a Tool for Monitoring Individual Proteins in Complex Mixtures.
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