Magic angle spinning NMR of proteins: high-frequency dynamic nuclear polarization and (1)H detection.

Abstract:

: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 (13)C- and (15)N-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 (1)H-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 10(2) to 10(3), thereby overcoming NMR's inherent low sensitivity. Alternatively, it permits structural investigations at the nanomolar scale. In addition, (1)H 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 (2)H-labeled proteins.

journal_name

Annu Rev Biochem

authors

Su Y,Andreas L,Griffin RG

doi

10.1146/annurev-biochem-060614-034206

subject

Has Abstract

pub_date

2015-01-01 00:00:00

pages

465-97

eissn

0066-4154

issn

1545-4509

journal_volume

84

pub_type

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