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Optimization of protein samples for NMR using thermal shift assays.

Kozak, Sandra
; 
Lercher, Lukas
; 
Karanth, Megha N
; 
Meijers, Rob
; ; 
Boivin, Stephane
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2016
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Abstract
Maintaining a stable fold for recombinant proteins is challenging, especially when working with highly purified and concentrated samples at temperatures >20 °C. Therefore, it is worthwhile to screen for different buffer components that can stabilize protein samples. Thermal shift assays or ThermoFluor(®) provide a high-throughput screening method to assess the thermal stability of a sample under several conditions simultaneously. Here, we describe a thermal shift assay that is designed to optimize conditions for nuclear magnetic resonance studies, which typically require stable samples at high concentration and ambient (or higher) temperature. We demonstrate that for two challenging proteins, the multicomponent screen helped to identify ingredients that increased protein stability, leading to clear improvements in the quality of the spectra. Thermal shift assays provide an economic and time-efficient method to find optimal conditions for NMR structural studies.
Citation
Optimization of protein samples for NMR using thermal shift assays. 2016, 64 (4):281-9 J. Biomol. NMR
Affiliation
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
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Journal
Journal of biomolecular NMR
URI
http://hdl.handle.net/10033/621280
DOI
10.1007/s10858-016-0027-z
PubMed ID
26984476
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Language
en
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ISSN
1573-5001
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publications of the research group NMR-based structural chemistry(NBSC)