2.50
Hdl Handle:
http://hdl.handle.net/10033/621280
Title:
Optimization of protein samples for NMR using thermal shift assays.
Authors:
Kozak, Sandra; Lercher, Lukas; Karanth, Megha N; Meijers, Rob; Carlomagno, Teresa ( 0000-0002-2437-2760 ) ; Boivin, Stephane
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.
Affiliation:
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
Optimization of protein samples for NMR using thermal shift assays. 2016, 64 (4):281-9 J. Biomol. NMR
Journal:
Journal of biomolecular NMR
Issue Date:
2016
URI:
http://hdl.handle.net/10033/621280
DOI:
10.1007/s10858-016-0027-z
PubMed ID:
26984476
Type:
Article
Language:
en
ISSN:
1573-5001
Appears in Collections:
publications of te research group NMR-based structural chemistry (NBSC)

Full metadata record

DC FieldValue Language
dc.contributor.authorKozak, Sandraen
dc.contributor.authorLercher, Lukasen
dc.contributor.authorKaranth, Megha Nen
dc.contributor.authorMeijers, Roben
dc.contributor.authorCarlomagno, Teresaen
dc.contributor.authorBoivin, Stephaneen
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.en
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

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