Journal Article

PUBDB-2021-00618

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png In‐depth interrogation of protein thermal unfolding data with MoltenProt

Kotov, V.CSSB*EMBL* ; Mlynek, G. ; Vesper, O.CSSB*DESY* ; Pletzer, M. ; Wald, J.CSSB* ; Teixeira-Duarte, C. M. ; Celia, H. ; Garcia-Alai, M. ; Nussberger, S. ; Buchanan, S. K. ; Morais-Cabral, J. H. ; Loew, C.CSSB*EMBL* ; Djinovic-Carugo, K. ; Marlovits, T. (Corresponding author)CSSB*

2021
Protein Society Bethesda, Md.

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Please use a persistent id in citations: doi:10.1002/pro.3986  doi:10.3204/PUBDB-2021-00618

Abstract: 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.

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Contributing Institute(s):

1. Centre for Structural Systems Biology (FS-CSSB-GS)
2. CSSB-EMBL-CL (CSSB-EMBL-CL)
3. CSSB-CF-SPC (CSSB-CF-SPC)
4. CSSB-UKE-TM (CSSB-UKE-TM)

Research Program(s):

1. 633 - Life Sciences – Building Blocks of Life: Structure and Function (POF4-633) (POF4-633)

Experiment(s):

1. No specific instrument

Appears in the scientific report 2021

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Database coverage:
; ; ; BIOSIS Previews ; Biological Abstracts ; Clarivate Analytics Master Journal List ; Current Contents - Life Sciences ; DEAL Wiley ; Essential Science Indicators ; IF < 5 ; JCR ; PubMed Central ; SCOPUS ; Science Citation Index Expanded ; Web of Science Core Collection

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