Sensitivity-Enhanced NMR Reveals Alterations in Protein Structure by Cellular Milieus

Kendra K. Frederick, Vladimir K. Michaelis, Björn Corzilius, Ta Chung Ong, Angela C. Jacavone, Robert G. Griffin, Susan Lindquist

Research output: Contribution to journalArticle

74 Scopus citations

Abstract

Summary Biological processes occur in complex environments containing a myriad of potential interactors. Unfortunately, limitations on the sensitivity of biophysical techniques normally restrict structural investigations to purified systems, at concentrations that are orders of magnitude above endogenous levels. Dynamic nuclear polarization (DNP) can dramatically enhance the sensitivity of nuclear magnetic resonance (NMR) spectroscopy and enable structural studies in biologically complex environments. Here, we applied DNP NMR to investigate the structure of a protein containing both an environmentally sensitive folding pathway and an intrinsically disordered region, the yeast prion protein Sup35. We added an exogenously prepared isotopically labeled protein to deuterated lysates, rendering the biological environment "invisible" and enabling highly efficient polarization transfer for DNP. In this environment, structural changes occurred in a region known to influence biological activity but intrinsically disordered in purified samples. Thus, DNP makes structural studies of proteins at endogenous levels in biological contexts possible, and such contexts can influence protein structure.

Original languageEnglish (US)
Pages (from-to)620-628
Number of pages9
JournalCell
Volume163
Issue number3
DOIs
StatePublished - Oct 22 2015

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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    Frederick, K. K., Michaelis, V. K., Corzilius, B., Ong, T. C., Jacavone, A. C., Griffin, R. G., & Lindquist, S. (2015). Sensitivity-Enhanced NMR Reveals Alterations in Protein Structure by Cellular Milieus. Cell, 163(3), 620-628. https://doi.org/10.1016/j.cell.2015.09.024