Gupta, R., et al., Dynamic Nuclear Polarization Enhanced MAS NMR Spectroscopy for Structural Analysis of HIV-1 Protein Assemblies. J Phys Chem B, 2016. 120(2): p. 329-39.
Mature infectious HIV-1 virions contain conical capsids composed of CA protein, generated by the proteolytic cleavage cascade of the Gag polyprotein, termed maturation. The mechanism of capsid core formation through the maturation process remains poorly understood. We present DNP-enhanced MAS NMR studies of tubular assemblies of CA and Gag CA-SP1 maturation intermediate and report 20-64-fold sensitivity enhancements due to DNP at 14.1 T. These sensitivity enhancements enabled direct observation of spacer peptide 1 (SP1) resonances in CA-SP1 by dipolar-based correlation experiments, unequivocally indicating that the SP1 peptide is unstructured in assembled CA-SP1 at cryogenic temperatures, corroborating our earlier results. Furthermore, the dependence of DNP enhancements and spectral resolution on magnetic field strength (9.4-18.8 T) and temperature (109-180 K) was investigated. Our results suggest that DNP-based measurements could potentially provide residue-specific dynamics information by allowing for the extraction of the temperature dependence of the anisotropic tensorial or relaxation parameters. With DNP, we were able to detect multiple well-resolved isoleucine side-chain conformers; unique intermolecular correlations across two CA molecules; and functionally relevant conformationally disordered states such as the 14-residue SP1 peptide, none of which are visible at ambient temperatures. The detection of isolated conformers and intermolecular correlations can provide crucial constraints for structure determination of these assemblies. Overall, our results establish DNP-based MAS NMR spectroscopy as an excellent tool for the characterization of HIV-1 assemblies.
Essentials of Dynamic Nuclear Polarization #DNPNMR
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Essentials of Dynamic Nuclear Polarization #DNPNMR
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05-25-2016 02:30 AM
Dynamic Nuclear Polarization enhanced NMR at 187GHz/284MHz using an Extended Interaction Klystron amplifier #DNPNMR
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Dynamic Nuclear Polarization enhanced NMR at 187GHz/284MHz using an Extended Interaction Klystron amplifier #DNPNMR
Kemp, T.F., et al., Dynamic Nuclear Polarization enhanced NMR at 187GHz/284MHz using an Extended Interaction Klystron amplifier. J Magn Reson, 2016. 265: p. 77-82.
http://www.ncbi.nlm.nih.gov/pubmed/26867091
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05-18-2016 09:53 PM
[NMR paper] Dynamic Nuclear Polarization Enhanced MAS NMR for Structural Analysis of HIV-1 Protein Assemblies.
Dynamic Nuclear Polarization Enhanced MAS NMR for Structural Analysis of HIV-1 Protein Assemblies.
Related Articles Dynamic Nuclear Polarization Enhanced MAS NMR for Structural Analysis of HIV-1 Protein Assemblies.
J Phys Chem B. 2015 Dec 28;
Authors: Gupta R, Lu M, Hou G, Caporini MA, Rosay M, Maas WE, Struppe JO, Suiter CL, Ahn J, Byeon IL, Franks WT, Orwick-Rydmark M, Bertarello A, Oschkinat H, Lesage A, Pintacuda G, Gronenborn AM, Polenova TE
Abstract
Mature infectious HIV-1 virions contain conical capsids comprised of CA...
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12-29-2015 08:05 PM
[NMR paper] NMR-based structural biology enhanced by dynamic nuclear polarization at high magnetic field.
NMR-based structural biology enhanced by dynamic nuclear polarization at high magnetic field.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--production.springer.de-OnlineResources-Logos-springerlink.gif Related Articles NMR-based structural biology enhanced by dynamic nuclear polarization at high magnetic field.
J Biomol NMR. 2014 Nov;60(2-3):157-68
Authors: Koers EJ, van der Cruijsen EA, Rosay M, Weingarth M, Prokofyev A, Sauvée C, Ouari O, van der Zwan J, Pongs O, Tordo P, Maas WE, Baldus M
Abstract
...
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06-24-2015 01:08 PM
NMR-based structural biology enhanced by dynamic nuclear polarization at high magnetic field
From The DNP-NMR Blog:
NMR-based structural biology enhanced by dynamic nuclear polarization at high magnetic field
Koers, E.J., et al., NMR-based structural biology enhanced by dynamic nuclear polarization at high magnetic field. J Biomol NMR, 2014. 60(2-3): p. 157-68.
http://www.ncbi.nlm.nih.gov/pubmed/25284462
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11-19-2014 04:32 PM
NMR-based structural biology enhanced by dynamic nuclear polarization at high magnetic field
NMR-based structural biology enhanced by dynamic nuclear polarization at high magnetic field
Abstract
Dynamic nuclear polarization (DNP) has become a powerful method to enhance spectroscopic sensitivity in the context of magnetic resonance imaging and nuclear magnetic resonance spectroscopy. We show that, compared to DNP at lower field (400Â*MHz/263Â*GHz), high field DNP (800Â*MHz/527Â*GHz) can significantly enhance spectral resolution and allows exploitation of the paramagnetic relaxation properties of DNP polarizing agents as direct structural...
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10-05-2014 09:01 AM
Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology
From The DNP-NMR Blog:
Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology
Gelis, I., et al., Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology. J. Biomol. NMR, 2013. 56(2): p. 85-93.
http://dx.doi.org/10.1007/s10858-013-9721-2
Dynamic Nuclear Polarization Enhanced MAS NMR Spectroscopy for Structural Analysis of HIV-1 Protein Assemblies #DNPNMR
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