Differential Dynamical Effects of Macromolecular Crowding on an Intrinsically Disordered Protein and a Globular Protein: Implications for In-Cell NMR Spectroscopy

In-cell NMR provides a valuable means to assess how macromolecules, with concentrations up to 300 g/L in the cytoplasm, affect the structure and dynamics of proteins at atomic resolution. Here an intrinsically disordered protein, α-synuclein (αSN), and a globular protein, chymotrypsin inhibitor 2 (C...

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Bibliographic Details
Published in:Journal of the American Chemical Society 2008-05, Vol.130 (20), p.6310-6311
Main Authors: Li, Conggang, Charlton, Lisa M, Lakkavaram, Asha, Seagle, Christopher, Wang, Guifang, Young, Gregory B, Macdonald, Jeffrey M, Pielak, Gary J
Format: Article
Language:English
Subjects:
alpha-Synuclein - chemistry
Nuclear Magnetic Resonance, Biomolecular - methods
Peptides - chemistry
Plant Proteins - chemistry
Povidone - chemistry
Viscosity
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Summary:In-cell NMR provides a valuable means to assess how macromolecules, with concentrations up to 300 g/L in the cytoplasm, affect the structure and dynamics of proteins at atomic resolution. Here an intrinsically disordered protein, α-synuclein (αSN), and a globular protein, chymotrypsin inhibitor 2 (CI2) were examined by using in-cell NMR. High-resolution in-cell spectra of αSN can be obtained, but CI2 leaks from the cell and the remaining intracellular CI2 is not detectable. Even after stabilizing the cells from leakage by using alginate encapsulation, no CI2 signal is detected. From in vitro studies we conclude that this difference in detectability is the result of the differential dynamical response of disordered and ordered proteins to the changes of motion caused by the increased viscosity in cells.
ISSN:0002-7863
1520-5126
DOI:10.1021/ja801020z