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Default In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.

In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.

Related Articles In-Cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.

Methods Mol Biol. 2020;2141:873-893

Authors: Gerez JA, Prymaczok NC, Riek R

Abstract
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 (?Syn) in mammalian cells. This chapter includes a simplified expression and purification protocol of recombinant ?Syn 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.


PMID: 32696394 [PubMed - in process]



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