Details
| Original language | English |
|---|---|
| Pages (from-to) | 281-289 |
| Number of pages | 9 |
| Journal | Journal of biomolecular NMR |
| Volume | 64 |
| Issue number | 4 |
| Early online date | 17 Mar 2016 |
| Publication status | Published - 1 Apr 2016 |
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.
Keywords
- Differential scanning fluorimetry, Nuclear magnetic resonance, Protein thermal stability, Sample optimization, Thermal shift assay, ThermoFluor
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Chemistry(all)
- Spectroscopy
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In: Journal of biomolecular NMR, Vol. 64, No. 4, 01.04.2016, p. 281-289.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Optimization of protein samples for NMR using thermal shift assays
AU - Kozak, Sandra
AU - Lercher, Lukas
AU - Karanth, Megha N.
AU - Meijers, Rob
AU - Carlomagno, Teresa
AU - Boivin, Stephane
PY - 2016/4/1
Y1 - 2016/4/1
N2 - 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.
AB - 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.
KW - Differential scanning fluorimetry
KW - Nuclear magnetic resonance
KW - Protein thermal stability
KW - Sample optimization
KW - Thermal shift assay
KW - ThermoFluor
UR - http://www.scopus.com/inward/record.url?scp=84961203813&partnerID=8YFLogxK
U2 - 10.1007/s10858-016-0027-z
DO - 10.1007/s10858-016-0027-z
M3 - Article
C2 - 26984476
AN - SCOPUS:84961203813
VL - 64
SP - 281
EP - 289
JO - Journal of biomolecular NMR
JF - Journal of biomolecular NMR
SN - 0925-2738
IS - 4
ER -