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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>
- 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<br />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.<br />Published by Protein Society, Bethesda, Md.
- Subjects :
- Protein Folding
Hot Temperature
Melting temperature
High-throughput screening
Crystallography, X-Ray
melting temperature
Biochemistry
high‐throughput screening
03 medical and health sciences
symbols.namesake
Thermal
Microscopy
ddc:610
MoltenProt
Molecular Biology
030304 developmental biology
Thermostability
0303 health sciences
Tools for Protein Science
buffer optimization
Chemistry
NanoDSF
protein unfolding
030302 biochemistry & molecular biology
Membrane Proteins
thermostability
Gibbs free energy
Membrane protein
Multiprotein Complexes
Unfolded protein response
symbols
Biological system
Software
Subjects
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