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In‐depth interrogation of protein thermal unfolding data with<scp>MoltenProt</scp>

In‐depth interrogation of protein thermal unfolding data with<scp>MoltenProt</scp>

Authors :
Herve Celia
Vadim Kotov
Oliver Vesper
Maria Garcia-Alai
Stephan Nussberger
Susan K. Buchanan
G. Mlynek
Jiri Wald
C. Loew
Celso M. Teixeira-Duarte
Marina Pletzer
João H. Morais-Cabral
Thomas C. Marlovits
Kristina Djinović-Carugo
Source :
Protein science 30(1), 201-217 (2021). doi:10.1002/pro.3986, Protein Science : A Publication of the Protein Society
Publication Year :
2020
Publisher :
Wiley, 2020.

Abstract

Protein science 30(1), 201 - 217 (2021). doi:10.1002/pro.3986&lt;br /&gt;Protein stability is a key factor in successful structural and biochemical research. However, the approaches for systematic comparison of protein stability are limited by sample consumption or compatibility with sample buffer components. Here we describe how miniaturized measurement of intrinsic tryptophan fluorescence (NanoDSF assay) in combination with a simplified description of protein unfolding can be used to interrogate the stability of a protein sample. We demonstrate that improved protein stability measures, such as apparent Gibbs free energy of unfolding, rather than melting temperature T$_m$, should be used to rank the results of thermostability screens. The assay is compatible with protein samples of any composition, including protein complexes and membrane proteins. Our data analysis software, MoltenProt, provides an easy and robust way to perform characterization of multiple samples. Potential applications of MoltenProt and NanoDSF include buffer and construct optimization for X‐ray crystallography and cryo‐electron microscopy, screening for small‐molecule binding partners and comparison of effects of point mutations.&lt;br /&gt;Published by Protein Society, Bethesda, Md.

Details

ISSN :
1469896X and 09618368
Volume :
30
Database :
OpenAIRE
Journal :
Protein Science
Accession number :
edsair.doi.dedup.....286fe9b9f64775e4766faec078c5cb02
Full Text :
https://doi.org/10.1002/pro.3986