## Targeted expression and purification of fluorine labelled cold shock protein B by using an auxotrophic strategy

2019
Journal article
Published
##### Published in
Protein expression and purification ; 157 (2019). - pp. 86-91. - ISSN 1046-5928. - eISSN 1096-0279
##### Abstract
High resolution NMR spectroscopy is a seminal method in modern structural biology to obtain insights into proteins' structure, dynamics and function at dilute condition as well as in a cell-like environment or even intracellularly. Usually, 1H, 15N or 13C nuclei are predominantly used for the characterization of the protein of interest. These measurements are limited due to the wealth of chemical shifts and background signals arising from all molecules present in the NMR test tube. On top of that, the protein under study has to be isotopically enriched in nitrogen and/or carbon nuclei enabling to overcome the inherently low natural abundance of 13C and 15N NMR active isotopes. In this way switching to 19F NMR spectroscopy strongly reduces the total amount of signals seen in an NMR spectrum as it turns off background signals and is for this reason extremely attractive for highly-resolved investigations of proteins performance measured directly in cells or in a cell-like environment. Here we show the effective expression and purification of cold shock protein B from Bacillus subtilis (BsCspB) using fluorine labelled phenylalanine or fluorine labelled tryptophan residues. We reveal that fluorine labelled BsCspB represents the same fold on a secondary as tertiary level as seen for the wild type protein independent of the labelling position illuminating the soft character of fluorine insertion. This experimental setup of targeted fluorine labelling sets a profound ground for a broad range of highly-resolved 19F NMR applications to be performed in a complex cellular environment.
540 Chemistry
##### Keywords
NMR spectroscopy, Cold shock protein B, E. coli, Recombinant protein expression, 19F labelling, Auxotrophy
##### Cite This
ISO 690WELTE, Hannah, Michael KOVERMANN, 2019. Targeted expression and purification of fluorine labelled cold shock protein B by using an auxotrophic strategy. In: Protein expression and purification. 157, pp. 86-91. ISSN 1046-5928. eISSN 1096-0279. Available under: doi: 10.1016/j.pep.2019.02.006
BibTex
@article{Welte2019-05Targe-45608,
year={2019},
doi={10.1016/j.pep.2019.02.006},
title={Targeted expression and purification of fluorine labelled cold shock protein B by using an auxotrophic strategy},
volume={157},
issn={1046-5928},
journal={Protein expression and purification},
pages={86--91},
author={Welte, Hannah and Kovermann, Michael}
}

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