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Methods Mol Biol. 2018;1688:133-154. doi: 10.1007/978-1-4939-7386-6_7.

Strategies for Efficient Sample Preparation for Dynamic Nuclear Polarization Solid-State NMR of Biological Macromolecules.

Author information

1
New York Structural Biology Center, 89 Convent Avenue, New York, NY, 10027, USA. bitin@nysbc.org.
2
Bruker Biospin Corporation, 15 Fortune Drive, Billerica, MA, 01821, USA.

Abstract

Solid-state NMR (SSNMR) is a powerful tool for the elucidation of structure and dynamics in biological macromolecules. Over the years, SSNMR spectroscopists have developed an array of techniques enabling the measurement of internuclear correlations, distances, and torsional angles; these have been applied to the study of a number of biological systems that are difficult to study by X-ray crystallography and solution NMR, including key biological targets such as membrane proteins and amyloid fibrils. Applications of SSNMR to other topic areas, including materials science, pharmaceuticals, and small molecules, have also flourished in recent years. These studies, however, have always been hampered by the low inherent sensitivity of SSNMR, requiring large amounts of both sample and time for data collection. By taking advantage of unpaired electrons doped into a sample as a ready source of additional nuclear polarization, dynamic nuclear polarization (DNP) has brought about large improvements in SSNMR sensitivity. These, in turn, have enabled structural studies of previously inaccessible targets, such as large protein complexes, nucleic acids, viral capsids, and membrane proteins in vivo. Herein, we focus on sample preparation strategies and considerations for scientists interested in applying DNP to challenging systems.

KEYWORDS:

Dynamic nuclear polarization; Magnetic resonance; Radicals; Signal enhancement; Solid-state NMR

PMID:
29151208
DOI:
10.1007/978-1-4939-7386-6_7
[Indexed for MEDLINE]

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