Category Archives: 1H ssNMR

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.

Dynamic nuclear polarization (DNP) is a powerful way to overcome the sensitivity limitation of magic?angle?spinning (MAS) NMR experiments. However, the resolution of the DNP?NMR spectra of proteins is compromised by severe line broadening associated with the necessity to perform experiments at cryogenic temperatures and in the presence of paramagnetic radicals. High?quality DNP?enhanced NMR spectra of the Acinetobacter phage 205 (AP205) nucleocapsid can be obtained by combining high magnetic field (800?MHz) and fast MAS (40?kHz). These conditions yield enhanced resolution and long coherence lifetimes allowing the acquisition of resolved 2D correlation spectra and of previously unfeasible scalar?based experiments. This enables the assignment of aromatic resonances of the AP205 coat protein and its packaged RNA, as well as the detection of long?range contacts, which are not observed at room temperature, opening new possibilities for structure determination.

Magic Angle Spinning NMR of Proteins: High-Frequency Dynamic Nuclear Polarization and H Detection

Su, Y., L. Andreas, and R.G. Griffin, Magic Angle Spinning NMR of Proteins: High-Frequency Dynamic Nuclear Polarization and H Detection. Annu Rev Biochem, 2015.

http://www.ncbi.nlm.nih.gov/pubmed/25839340

Magic angle spinning (MAS) NMR studies of amyloid and membrane proteins and large macromolecular complexes are an important new approach to structural biology. However, the applicability of these experiments, which are based on 13C- and 15N-detected spectra, would be enhanced if the sensitivity were improved. Here we discuss two advances that address this problem: high-frequency dynamic nuclear polarization (DNP) and 1H-detected MAS techniques. DNP is a sensitivity enhancement technique that transfers the high polarization of exogenous unpaired electrons to nuclear spins via microwave irradiation of electron-nuclear transitions. DNP boosts NMR signal intensities by factors of 102 to 103, thereby overcoming NMR’s inherent low sensitivity. Alternatively, it permits structural investigations at the nanomolar scale. In addition, 1H detection is feasible primarily because of the development of MAS rotors that spin at frequencies of 40 to 60 kHz or higher and the preparation of extensively 2H-labeled proteins. Expected final online publication date for the Annual Review of Biochemistry Volume 84 is June 02, 2015. Please see http://www.annualreviews.org/catalog/pubdates.aspx for revised estimates.

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