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Solid-state NMR of a protein in a precipitated complex with a full-length antibody

Lamley, Jonathan M. and Iuga, Dinu and Oster, Carl and Sass, Hans Juergen and Rogowski, Marco and Oss, Andres and Past, Jaan and Reinhold, Andres and Grzesiek, Stephan and 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, Vol. 136, H. 48. pp. 16800-16806.

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Official URL: http://edoc.unibas.ch/dok/A6328781

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Abstract

NMR 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 application of (1)H-detected experiments at <50 kHz magic angle spinning frequencies 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 hundreds-of-kDa protein complexes. Protein-protein interaction interfaces can be mapped out by comparing the chemical shifts of proteins within solid-state complexes with those of the same constituent proteins free in solution. We employ this methodology to characterize a <300 kDa complex of GB1 with full-length human immunoglobulin, where we find 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 investigate extensions of our methodology to spinning frequencies of up to 100 kHz.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Grzesiek)
UniBasel Contributors:Grzesiek, Stephan and Rogowski, Marco and Sass, Hans-Jürgen
Item Type:Article, refereed
Article Subtype:Research Article
Bibsysno:Link to catalogue
Publisher:American Chemical Society
ISSN:0002-7863
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:09 Jan 2015 09:25
Deposited On:09 Jan 2015 09:25

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