Kim, Yaewon, Mengxiao Liu, and Christian Hilty. “Determination of Binding Affinities Using Hyperpolarized NMR with Simultaneous 4-Channel Detection.” Journal of Magnetic Resonance 295 (October 2018): 80–86.
Dissolution dynamic nuclear polarization (D-DNP) is a powerful technique to improve NMR sensitivity by a factor of thousands. Combining D-DNP with NMR-based screening enables to mitigate solubility or availability problems of ligands and target proteins in drug discovery as it can lower the concentration requirements into the sub-micromolar range. One of the challenges that D-DNP assisted NMR screening methods face for broad application, however, is a reduced throughput due to additional procedures and time required to create hyperpolarization. These requirements result in a delay of several tens of minutes in-between each NMR measurement. To solve this problem, we have developed a simultaneous 4-channel detection method for hyperpolarized 19F NMR, which can increase throughput four-fold utilizing a purpose-built multiplexed NMR spectrometer and probe. With this system, the concentration-dependent binding interactions were observed for benzamidine and benzylamine with the serine protease trypsin. A T2 relaxation measurement of a hyperpolarized reporter ligand (TFBC; CF3C6H4CNHNH2), which competes for the same binding site on the trypsin with the other ligands, was used. The hyperpolarized TFBC was mixed with trypsin and the ligand of interest, and injected into four flow cells inside the NMR probe. Across the set of four channels, a concentration gradient was created. From the simultaneously acquired relaxation datasets, it was possible to determine the dissociation constant (KD) of benzamidine or benzylamine without the requirement for individually optimizing experimental conditions for different affinities. A simulation showed that this 4-channel detection method applied to D-DNP NMR extends the screenable KD range to up to three orders of magnitude in a single experiment.
Simultaneous Cross Polarization to 13C and 15N with 1H Detection at 60 kHz MAS Solid-state NMR
Simultaneous Cross Polarization to 13C and 15N with 1H Detection at 60 kHz MAS Solid-state NMR
Publication date: Available online 12 December 2015
Source:Journal of Magnetic Resonance</br>
Author(s): Bibhuti B. Das, Stanley J. Opella</br>
We describe high resolution MAS solid-state NMR experiments that utilize 1H detection with 60 kHz magic angle spinning; simultaneous cross-polarization from 1H to 15N and 13C nuclei; bidirectional cross-polarization between 13C and 15N nuclei; detection of both amide nitrogen and aliphatic carbon 1H; and measurement of both...
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12-12-2015 05:40 PM
Simultaneous hyperpolarized (13)C-pyruvate MRI and (18)F-FDG-PET in cancer (hyperPET): feasibility of a new imaging concept using a clinical PET/MRI scanner
From The DNP-NMR Blog:
Simultaneous hyperpolarized (13)C-pyruvate MRI and (18)F-FDG-PET in cancer (hyperPET): feasibility of a new imaging concept using a clinical PET/MRI scanner
Gutte, H., et al., Simultaneous hyperpolarized (13)C-pyruvate MRI and (18)F-FDG-PET in cancer (hyperPET): feasibility of a new imaging concept using a clinical PET/MRI scanner. American Journal of Nuclear Medicine and Molecular Imaging, 2015. 5(1): p. 38-45.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4299777/
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05-14-2015 04:52 AM
O-tert-Butyltyrosine, an NMR Tag for High-Molecular-Weight Systems and Measurements of Submicromolar Ligand Binding Affinities
O-tert-Butyltyrosine, an NMR Tag for High-Molecular-Weight Systems and Measurements of Submicromolar Ligand Binding Affinities
Wan-Na Chen, Kekini Vahini Kuppan, Michael David Lee, Kristaps Jaudzems, Thomas Huber and Gottfried Otting
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/jacs.5b01918/20150327/images/medium/ja-2015-01918z_0002.gif
Journal of the American Chemical Society
DOI: 10.1021/jacs.5b01918
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA ...
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03-27-2015 11:59 PM
[NMR paper] O-tert-Butyltyrosine, an NMR tag for high-molecular weight systems and measurements of submicromolar ligand binding affinities.
O-tert-Butyltyrosine, an NMR tag for high-molecular weight systems and measurements of submicromolar ligand binding affinities.
O-tert-Butyltyrosine, an NMR tag for high-molecular weight systems and measurements of submicromolar ligand binding affinities.
J Am Chem Soc. 2015 Mar 19;
Authors: Chen WN, Kuppan KV, Lee MD, Jaudzems K, Huber T, Otting G
Abstract
O-tert-butyl-tyrosine (Tby) is an unnatural amino acid that can be site-specifically incorporated into proteins using established orthogonal aminoacyl-tRNA synthetase/tRNA...
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03-20-2015 04:15 PM
[NMR paper] Identification of differential protein binding affinities in an atropisomeric pharmaceutical compound using non-covalent mass spectrometry, equilibrium dialysis and NMR.
Identification of differential protein binding affinities in an atropisomeric pharmaceutical compound using non-covalent mass spectrometry, equilibrium dialysis and NMR.
Related Articles Identification of differential protein binding affinities in an atropisomeric pharmaceutical compound using non-covalent mass spectrometry, equilibrium dialysis and NMR.
Anal Chem. 2013 May 22;
Authors: Maple HJ, Garlish RA, Whitcombe I, Hold A, Prosser CE, Ford D, Mackenzie H, Crosby J, Porter J, Taylor RJ, Crump MP
Abstract
Atropisomerism of...
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05-24-2013 10:44 PM
Simultaneous Detection of Protein Phosphorylation and Acetylation by High-Resolution
Simultaneous Detection of Protein Phosphorylation and Acetylation by High-Resolution NMR Spectroscopy.
Simultaneous Detection of Protein Phosphorylation and Acetylation by High-Resolution NMR Spectroscopy.
J Am Chem Soc. 2010 Oct 1;
Authors: Liokatis S, Dose A, Schwarzer D, Selenko P
Post-translational protein modifications (PTMs) such as phosphorylation and acetylation regulate a large number of eukaryotic signaling processes. In most instances, it is the combination of different PTMs that "encode" the biological outcome of these covalent...
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10-05-2010 12:11 PM
Simultaneous Detection of Protein Phosphorylation and Acetylation by High-Resolution
Simultaneous Detection of Protein Phosphorylation and Acetylation by High-Resolution NMR Spectroscopy
Stamatios Liokatis, Alexander Dose, Dirk Schwarzer and Philipp Selenko
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja106764y/aop/images/medium/ja-2010-06764y_0002.gif
Journal of the American Chemical Society
DOI: 10.1021/ja106764y
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/jfhp2Sg-hpY
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10-02-2010 12:16 AM
Simultaneous detection of amide and methyl correlations using a time shared NMR experiment: application to binding epitope mapping
Simultaneous detection of amide and methyl correlations using a time shared NMR experiment: application to binding epitope mapping
Peter Würtz, Olli Aitio, Maarit Hellman and Perttu Permi
Journal of Biomolecular NMR; 2007; 39(2) pp 97 - 105
Abstract:
Simultaneous recording of different NMR parameters is an efficient way to reduce the overall experimental time and speed up structural studies of biological macromolecules. This can especially be beneficial in the case of fast NMR-based drug screening applications or for collecting NOE restraints, where prohibitively long data collection...