High resolution NMR spectroscopy of nanocrystalline proteins at ultra-high magnetic field

Magic-angle spinning (MAS) solid-state NMR (SSNMR) spectroscopy of uniformly-¹³C,¹⁵N labeled protein samples provides insight into atomic-resolution chemistry and structure. Data collection efficiency has advanced remarkably in the last decade; however, the study of larger proteins is still challeng...

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Bibliographic Details
Published in:Journal of biomolecular NMR 2010-02, Vol.46 (2), p.149-155
Main Authors: Sperling, Lindsay J, Nieuwkoop, Andrew J, Lipton, Andrew S, Berthold, Deborah A, Rienstra, Chad M
Format: Article
Language:English
Subjects:
Biochemistry
Biological and Medical Physics
Biophysics
Crystallography, X-Ray
Data collection
Escherichia coli - enzymology
Escherichia coli Proteins - chemistry
Magnetic fields
Magnetics
Methylation
Nanoparticles - chemistry
Nerve Tissue Proteins - chemistry
Nuclear Magnetic Resonance, Biomolecular - methods
Physics
Physics and Astronomy
Protein Disulfide-Isomerases - chemistry
Protein Structure, Tertiary
Spectroscopy
Spectroscopy/Spectrometry
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Summary:Magic-angle spinning (MAS) solid-state NMR (SSNMR) spectroscopy of uniformly-¹³C,¹⁵N labeled protein samples provides insight into atomic-resolution chemistry and structure. Data collection efficiency has advanced remarkably in the last decade; however, the study of larger proteins is still challenged by relatively low resolution in comparison to solution NMR. In this study, we present a systematic analysis of SSNMR protein spectra acquired at 11.7, 17.6 and 21.1 Tesla (¹H frequencies of 500, 750, and 900 MHz). For two protein systems—GB1, a 6 kDa nanocrystalline protein and DsbA, a 21 kDa nanocrystalline protein—line narrowing is demonstrated in all spectral regions with increasing field. Resolution enhancement is greatest in the aliphatic region, including methine, methylene and methyl sites. The resolution for GB1 increases markedly as a function of field, and for DsbA, resolution in the C-C region increases by 42%, according to the number of peaks that can be uniquely picked and integrated in the 900 MHz spectra when compared to the 500 MHz spectra. Additionally, chemical exchange is uniquely observed in the highest field spectra for at least two isoleucine Cδ1 sites in DsbA. These results further illustrate the benefits of high-field MAS SSNMR spectroscopy for protein structural studies.
ISSN:0925-2738
1573-5001
DOI:10.1007/s10858-009-9389-9