Journal article
In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.
- Gerez JA Laboratory of Physical Chemistry, Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, Switzerland. juan.gerez@phys.chem.ethz.ch.
- Prymaczok NC Laboratory of Physical Chemistry, Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, Switzerland.
- Riek R Laboratory of Physical Chemistry, Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, Switzerland. roland.riek@phys.chem.ethz.ch.
- 2020-07-23
Published in:
- Methods in molecular biology (Clifton, N.J.). - 2020
English
In-cell NMR enables structural insights at atomic resolution of proteins in their natural environment. To date, very few methods have been developed to study proteins by in-cell NMR in mammalian systems. Here we describe a detailed protocol to conduct in-cell NMR on the intrinsically disordered protein of alpha-Synuclein (αSyn) in mammalian cells. This chapter includes a simplified expression and purification protocol of recombinant αSyn and its delivery into mammalian cells. The chapter also describes how to assess the cell leakage that might occur to the cells, the setup of the instrument, and how to perform basic analyses with the obtained NMR data.
- Language
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- English
- Open access status
- closed
- Identifiers
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- DOI 10.1007/978-1-0716-0524-0_45
- PMID 32696394
- Persistent URL
- https://sonar.ch/global/documents/77869
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