Sensitivity-Enhanced NMR Reveals Alterations in Protein Structure by Cellular Milieus
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Griffin_Sensitivity-enhanced.pdf
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Author(s)
Frederick, Kendra K.
Michaelis, Vladimir K.
Corzilius, Bjorn
Ong, Ta-Chung
Jacavone, Angela
Griffin, Robert Guy
Lindquist, Susan
Date Issued
October 2015
Journal
Cell
Publisher
Elsevier
Citation
Frederick, Kendra K.; Michaelis, Vladimir K.; Corzilius, Björn; Ong, Ta-Chung; Jacavone, Angela C.; Griffin, Robert G. and Lindquist, Susan. “Sensitivity-Enhanced NMR Reveals Alterations in Protein Structure by Cellular Milieus.” Cell 163, no. 3 (October 2015): 620–628. © 2015 Elsevier Inc
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Author's final manuscript
Abstract
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.
MIT Department
Massachusetts Institute of Technology. Department of Chemistry
Francis Bitter Magnet Laboratory (Massachusetts Institute of Technology)
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Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/j.cell.2015.09.024
