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Solid-state NMR of a protein in a precipitated complex with a full-length antibody
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Lamley, Jonathan M., Iuga, Dinu, Öster, Carl, Sass, Hans Jürgen, Rogowski, Marco, Oss, Andres, Past, Jaan, Reinhold, Andres, Grzesiek, Stephan, Samoson, Ago and Lewandowski, Józef R. (2014) Solid-state NMR of a protein in a precipitated complex with a full-length antibody. Journal of the American Chemical Society, Volume 136 (Number 48). pp. 16800-16806. doi:10.1021/ja5069992 ISSN 0002-7863.
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Official URL: http://dx.doi.org/10.1021/ja5069992
Abstract
NMR spectroscopy is a prime technique for characterizing atomic-resolution structures and dynamics of biomolecular complexes but for such systems faces challenges of sensitivity and spectral resolution. We demonstrate that the application of 1H-detected experiments at magic-angle spinning frequencies of >50 kHz enables the recording, in a matter of minutes to hours, of solid-state NMR spectra suitable for quantitative analysis of protein complexes present in quantities as small as a few nanomoles (tens of micrograms for the observed component). This approach enables direct structure determination and quantitative dynamics measurements in domains of protein complexes with masses of hundreds of kilodaltons. Protein–protein interaction interfaces can be mapped out by comparison of the chemical shifts of proteins within solid-state complexes with those of the same constituent proteins free in solution. We employed this methodology to characterize a >300 kDa complex of GB1 with full-length human immunoglobulin, where we found that sample preparation by simple precipitation yields spectra of exceptional quality, a feature that is likely to be shared with some other precipitating complexes. Finally, we investigated extensions of our methodology to spinning frequencies of up to 100 kHz.
Item Type: | Journal Article | ||||||
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Subjects: | Q Science > QC Physics Q Science > QD Chemistry |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry Faculty of Science, Engineering and Medicine > Science > Physics |
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Library of Congress Subject Headings (LCSH): | Nuclear magnetic resonance spectroscopy, Complex compounds -- Spectra | ||||||
Journal or Publication Title: | Journal of the American Chemical Society | ||||||
Publisher: | American Chemical Society | ||||||
ISSN: | 0002-7863 | ||||||
Official Date: | 10 October 2014 | ||||||
Dates: |
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Volume: | Volume 136 | ||||||
Number: | Number 48 | ||||||
Number of Pages: | 7 | ||||||
Page Range: | pp. 16800-16806 | ||||||
DOI: | 10.1021/ja5069992 | ||||||
Status: | Peer Reviewed | ||||||
Publication Status: | Published | ||||||
Access rights to Published version: | Restricted or Subscription Access | ||||||
Date of first compliant deposit: | 29 December 2015 | ||||||
Date of first compliant Open Access: | 29 December 2015 | ||||||
Funder: | Royal Society (Great Britain), Engineering and Physical Sciences Research Council (EPSRC), Estonian Science Foundation (ESF), Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung [Swiss National Science Foundation] (SNSF), Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), University of Warwick, Birmingham Science City, Advantage West Midlands (AWM), European Regional Development Fund (ERDF) | ||||||
Grant number: | RG130022 (RS), EP/L025906/1 (EPSRC), 31-109712 (SNSF), 31-132857 (SNSF) |
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