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dc.contributor.authorKozak, Sandra
dc.contributor.authorLercher, Lukas
dc.contributor.authorKaranth, Megha N
dc.contributor.authorMeijers, Rob
dc.contributor.authorCarlomagno, Teresa
dc.contributor.authorBoivin, Stephane
dc.date.accessioned2018-02-15T13:50:18Z
dc.date.available2018-02-15T13:50:18Z
dc.date.issued2016
dc.identifier.citationOptimization of protein samples for NMR using thermal shift assays. 2016, 64 (4):281-9 J. Biomol. NMRen
dc.identifier.issn1573-5001
dc.identifier.pmid26984476
dc.identifier.doi10.1007/s10858-016-0027-z
dc.identifier.urihttp://hdl.handle.net/10033/621280
dc.description.abstractMaintaining 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.
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subject.meshFluorometryen
dc.subject.meshMagnetic Resonance Spectroscopyen
dc.subject.meshNuclear Magnetic Resonance, Biomolecularen
dc.subject.meshProtein Stabilityen
dc.subject.meshProteinsen
dc.subject.meshTemperatureen
dc.titleOptimization of protein samples for NMR using thermal shift assays.en
dc.typeArticleen
dc.contributor.departmentHelmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalJournal of biomolecular NMRen
refterms.dateFOA2018-06-12T23:17:09Z
html.description.abstractMaintaining 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.


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