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Insights into Protein Stability in Cell Lysate by 19 F NMR Spectroscopy
Author(s) -
Welte Hannah,
Kovermann Michael
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202000413
Subject(s) - lysis , macromolecule , bacillus subtilis , nuclear magnetic resonance spectroscopy , protein stability , intracellular , macromolecular crowding , biophysics , protein folding , function (biology) , folding (dsp implementation) , chemistry , protein function , protein dynamics , biochemistry , protein structure , biology , microbiology and biotechnology , bacteria , stereochemistry , gene , electrical engineering , genetics , engineering
In living organisms, protein folding and function take place in an inhomogeneous, highly crowded environment possessing a concentration of diverse macromolecules of up to 400 g/L. It has been shown that the intracellular environment has a pronounced effect on the stability, dynamics and function of the protein under study, and has for this reason to be considered. However, most protein studies neglect the presence of these macromolecules. Consequently, we probe here the overall thermodynamic stability of cold shock protein B from Bacillus subtilis ( Bs CspB) in cell lysate. We found that an increase in cell lysate concentration causes a monotonic increase in the thermodynamic stability of Bs CspB. This result strongly underlines the importance of considering the biological environment when inherent protein parameters are quantitatively determined. Moreover, we demonstrate that targeted application of 19 F NMR spectroscopy operates as an ideal tool for protein studies performed in complex cellular surroundings.