Publication date: Available online 8 October 2015 Source:Cell
Author(s): Kendra*K. Frederick, Vladimir*K. Michaelis, Björn Corzilius, Ta-Chung Ong, Angela*C. Jacavone, Robert*G. Griffin, Susan Lindquist
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. Graphical abstract
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Sensitivity-enhanced NMR enabling structural analysis of protein at endogenous levels in a native biological context reveals that the cellular environment alters the structure of an intrinsically disordered protein domain.
HyperBIRD: A Sensitivity-Enhanced Approach to Collecting Homonuclear-Decoupled Proton NMR Spectra
From The DNP-NMR Blog:
HyperBIRD: A Sensitivity-Enhanced Approach to Collecting Homonuclear-Decoupled Proton NMR Spectra
Donovan, K.J. and L. Frydman, HyperBIRD: A Sensitivity-Enhanced Approach to Collecting Homonuclear-Decoupled Proton NMR Spectra. Angew Chem Int Ed Engl, 2014: p. n/a-n/a.
http://www.ncbi.nlm.nih.gov/pubmed/25256418
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Optimization of an absolute sensitivity in a glassy matrix during DNP-enhanced multidimensional solid-state NMR experiments
From The DNP-NMR Blog:
Optimization of an absolute sensitivity in a glassy matrix during DNP-enhanced multidimensional solid-state NMR experiments
Takahashi, H., et al., Optimization of an absolute sensitivity in a glassy matrix during DNP-enhanced multidimensional solid-state NMR experiments. J Magn Reson, 2013. 239C(0): p. 91-99.
http://www.ncbi.nlm.nih.gov/pubmed/24480716
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From The DNP-NMR Blog:
Sensitivity-enhanced solid-state NMR detection of expansin's target in plant cell walls
Wang, T., et al., Sensitivity-enhanced solid-state NMR detection of expansin's target in plant cell walls. Proc Natl Acad Sci U S A, 2013. 110(41): p. 16444-9.
http://www.ncbi.nlm.nih.gov/pubmed/24065828
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[NMR paper] Sensitivity-enhanced solid-state NMR detection of expansin's target in plant cell walls.
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Proc Natl Acad Sci U S A. 2013 Sep 24;
Authors: Wang T, Park YB, Caporini MA, Rosay M, Zhong L, Cosgrove DJ, Hong M
Abstract
Structure determination of protein binding to noncrystalline macromolecular assemblies such as plant cell walls (CWs) poses a significant structural biology challenge. CWs are loosened during growth by expansin proteins,...
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Acc Chem Res. 2013 Jul 26;
Authors: Parthasarathy S, Nishiyama Y, Ishii Y
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Recent research in fast magic angle spinning (MAS) methods has drasticallyimproved the resolution and sensitivity of NMR spectroscopy of biomolecules and materials in solids. In...
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Authors: Hong M, Yamaguchi S
A method for enhancing the sensitivity of 15N spectra of nonspinning solids through 1H indirect detection is introduced. By sampling the 1H signals in the windows of a pulsed spin-lock sequence, high-sensitivity 1H spectra can be obtained in two-dimensional (2D) spectra whose indirect dimension yields the 15N chemical...
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[NMR paper] Sensitivity enhanced NMR spectroscopy by quenching scalar coupling mediated relaxatio
Sensitivity enhanced NMR spectroscopy by quenching scalar coupling mediated relaxation: application to the direct observation of hydrogen bonds in 13C/15N-labeled proteins.
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J Biomol NMR. 2000 May;17(1):55-61
Authors: Liu A, Hu W, Qamar S, Majumdar A
In this paper, we demonstrate that the sensitivity of triple-resonance NMR experiments can be enhanced...
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Structure of key protein for cellular signal transduction elucidated - News-Medical.n
Structure of key protein for cellular signal transduction elucidated - News-Medical.net
<img alt="" height="1" width="1" />
Structure of key protein for cellular signal transduction elucidated
News-Medical.net
Using NMR spectroscopy, Professor Michael Sattler and his team elucidated the spatial structure of the Qua1 region of Sam68, which is responsible for the ...
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Sensitivity-Enhanced NMR Reveals Alterations in Protein Structure by Cellular Milieus
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