Related ArticlesSelective high-resolution DNP-enhanced NMR of biomolecular binding sites.
Chem Sci. 2019 Mar 21;10(11):3366-3374
Authors: Marin-Montesinos I, Goyard D, Gillon E, Renaudet O, Imberty A, Hediger S, De Paëpe G
Abstract
Locating binding sites in biomolecular assemblies and solving their structures are of the utmost importance to unravel functional aspects of the system and provide experimental data that can be used for structure-based drug design. This often still remains a challenge, both in terms of selectivity and sensitivity for X-ray crystallography, cryo-electron microscopy and NMR. In this work, we introduce a novel method called Selective Dynamic Nuclear Polarization (Sel-DNP) that allows selective highlighting and identification of residues present in the binding site. This powerful site-directed approach relies on the use of localized paramagnetic relaxation enhancement induced by a ligand-functionalized paramagnetic construct combined with difference spectroscopy to recover high-resolution and high-sensitivity information from binding sites. The identification of residues involved in the binding is performed using spectral fingerprints obtained from a set of high-resolution multidimensional spectra with varying selectivities. The methodology is demonstrated on the galactophilic lectin LecA, for which we report well-resolved DNP-enhanced spectra with linewidths between 0.5 and 1 ppm, which enable the de novo assignment of the binding interface residues, without using previous knowledge of the binding site location. Since this approach produces clean and resolved difference spectra containing a limited number of residues, resonance assignment can be performed without any limitation with respect to the size of the biomolecular system and only requires the production of one protein sample (e.g. 13C,15N-labeled protein).
Selective high-resolution DNP-enhanced NMR of biomolecular binding sites #DNPNMR
From The DNP-NMR Blog:
Selective high-resolution DNP-enhanced NMR of biomolecular binding sites #DNPNMR
Marin-Montesinos, Ildefonso, David Goyard, Emilie Gillon, Olivier Renaudet, Anne Imberty, Sabine Hediger, and Gaël De Paëpe. “Selective High-Resolution DNP-Enhanced NMR of Biomolecular Binding Sites.” Chemical Science 10, no. 11 (2019): 3366–74.
https://doi.org/10.1039/C8SC05696J.
nmrlearner
News from NMR blogs
0
03-28-2019 01:56 AM
Dynamic Nuclear Polarization-Enhanced Biomolecular NMR Spectroscopy at High Magnetic Field with Fast Magic-Angle Spinning #DNPNMR
From The DNP-NMR Blog:
Dynamic Nuclear Polarization-Enhanced Biomolecular NMR Spectroscopy at High Magnetic Field with Fast Magic-Angle Spinning #DNPNMR
Jaudzems, Kristaps, Andrea Bertarello, Sachin R. Chaudhari, Andrea Pica, Diane Cala-De Paepe, Emeline Barbet-Massin, Andrew J. Pell, et al. “Dynamic Nuclear Polarization-Enhanced Biomolecular NMR Spectroscopy at High Magnetic Field with Fast Magic-Angle Spinning.” Angewandte Chemie 0 (2018).
https://doi.org/10.1002/ange.201801016.
nmrlearner
News from NMR blogs
0
07-06-2018 09:40 AM
[NMR paper] Dynamic nuclear polarization enhanced biomolecular NMR spectroscopy at high magnetic field with fast magic-angle spinning.
Dynamic nuclear polarization enhanced biomolecular NMR spectroscopy at high magnetic field with fast magic-angle spinning.
Dynamic nuclear polarization enhanced biomolecular NMR spectroscopy at high magnetic field with fast magic-angle spinning.
Angew Chem Int Ed Engl. 2018 Mar 22;:
Authors: Jaudzems K, Bertarello A, Chaudhari SR, Pica A, Cala-De Paepe D, Barbet-Massin E, Pell AJ, Akopjana I, Kotelovica S, Gajan D, Ouari O, Tars K, Pintacuda G, Lesage A
Abstract
Dynamic nuclear polarization (DNP) represents a powerful way to...
nmrlearner
Journal club
0
03-23-2018 11:18 AM
[NMR paper] High-resolution 2D NMR of disordered proteins enhanced by hyperpolarized water.
High-resolution 2D NMR of disordered proteins enhanced by hyperpolarized water.
High-resolution 2D NMR of disordered proteins enhanced by hyperpolarized water.
Anal Chem. 2018 Mar 12;:
Authors: Szekely O, Olsen GL, Felli IC, Frydman L
Abstract
This study demonstrates the usefulness derived from relying on hyperpolarized water obtained by dissolution DNP, for site-resolved biophysical NMR studies of intrinsically disordered proteins. Thanks to the facile amide-solvent exchange experienced by protons in these proteins, 2D NMR...
nmrlearner
Journal club
0
03-13-2018 07:39 PM
[NMR paper] The effect of drug binding on specific sites in transmembrane helices 4 and 6 of the ABC exporter MsbA studied by DNP-enhanced solid-state NMR.
The effect of drug binding on specific sites in transmembrane helices 4 and 6 of the ABC exporter MsbA studied by DNP-enhanced solid-state NMR.
Related Articles The effect of drug binding on specific sites in transmembrane helices 4 and 6 of the ABC exporter MsbA studied by DNP-enhanced solid-state NMR.
Biochim Biophys Acta. 2017 Oct 22;:
Authors: Spadaccini R, Kaur H, Becker-Baldus J, Glaubitz C
Abstract
MsbA, a homodimeric ABC exporter, translocates its native substrate lipid A as well as a range of smaller, amphiphilic...
nmrlearner
Journal club
0
10-28-2017 08:03 AM
[NMR paper] Identification of heteromolecular binding sites in transcription factors Sp1 and TAF4 using high-resolution NMR spectroscopy.
Identification of heteromolecular binding sites in transcription factors Sp1 and TAF4 using high-resolution NMR spectroscopy.
Related Articles Identification of heteromolecular binding sites in transcription factors Sp1 and TAF4 using high-resolution NMR spectroscopy.
Protein Sci. 2017 Aug 31;:
Authors: Hibino E, Inoue R, Sugiyama M, Kuwahara J, Matsuzaki K, Hoshino M
Abstract
The expression of eukaryotic genes is precisely controlled by interactions between general transcriptional factors and promoter-specific transcriptional...
nmrlearner
Journal club
0
09-02-2017 02:29 AM
Identification of heteromolecular binding sites in transcription factors Sp1 and TAF4 using high-resolution NMR spectroscopy
Identification of heteromolecular binding sites in transcription factors Sp1 and TAF4 using high-resolution NMR spectroscopy
Abstract
The expression of eukaryotic genes is precisely controlled by interactions between general transcriptional factors and promoter-specific transcriptional activators.
The fourth element of TATA-box binding protein-associated factor (TAF4), an essential subunit of the general transcription factor TFIID, serves as a coactivator for various promoter-specific transcriptional regulators. Interactions between TAF4 and site-specific transcriptional...
nmrlearner
Journal club
0
08-31-2017 01:22 PM
High resolution methyl selective 13C-NMR of proteins in solution and solid state
High resolution methyl selective 13C-NMR of proteins in solution and solid state
Abstract New 13C-detected NMR experiments have been devised for molecules in solution and solid state, which provide chemical shift correlations of methyl groups with high resolution, selectivity and sensitivity. The experiments achieve selective methyl detection by exploiting the one bond J-coupling between the 13C-methyl nucleus and its directly attached 13C spin in a molecule. In proteins such correlations edit the 13C-resonances of different methyl containing residues into distinct spectral regions...
Selective high-resolution DNP-enhanced NMR of biomolecular binding sites.
Linear Mode
