MicroScale Thermophoresis

• Verfasst von: Jerabek-Willemsen, Moran [VerfasserIn]
• Verfasst von: André, Timon [VerfasserIn]
• Titel: MicroScale Thermophoresis
• Titelzusatz: interaction analysis and beyond
• Verf.angabe: Moran Jerabek-Willemsen, Timon André, Randy Wanner, Heide Marie Roth, Stefan Duhr, Philipp Baaske, Dennis Breitsprecher
• E-Jahr: 2014
• Jahr: 16 March 2014
• Umfang: 13 S.
• Fussnoten: Gesehen am 22.10.2020
• Titel Quelle: Enthalten in: Journal of molecular structure
• Ort Quelle: New York, NY [u.a.] : Elsevier, 1967
• Jahr Quelle: 2014
• Band/Heft Quelle: 1077(2014), Seite 101-113
• ISSN Quelle: 1872-8014
• DOI: doi:10.1016/j.molstruc.2014.03.009
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Binding studies
• Sach-SW: Interaction analysis
• Sach-SW: ITC
• Sach-SW: Microscale Thermophoresis
• Sach-SW: Protein folding
• Sach-SW: SPR
• K10plus-PPN: 1736242687

Abstract

MicroScale Thermophoresis (MST) is a powerful technique to quantify biomolecular interactions. It is based on thermophoresis, the directed movement of molecules in a temperature gradient, which strongly depends on a variety of molecular properties such as size, charge, hydration shell or conformation. Thus, this technique is highly sensitive to virtually any change in molecular properties, allowing for a precise quantification of molecular events independent of the size or nature of the investigated specimen. During a MST experiment, a temperature gradient is induced by an infrared laser. The directed movement of molecules through the temperature gradient is detected and quantified using either covalently attached or intrinsic fluorophores. By combining the precision of fluorescence detection with the variability and sensitivity of thermophoresis, MST provides a flexible, robust and fast way to dissect molecular interactions. In this review, we present recent progress and developments in MST technology and focus on MST applications beyond standard biomolecular interaction studies. By using different model systems, we introduce alternative MST applications - such as determination of binding stoichiometries and binding modes, analysis of protein unfolding, thermodynamics and enzyme kinetics. In addition, wedemonstrate the capability of MST to quantify high-affinity interactions with dissociation constants (Kds) in the low picomolar (pM) range as well as protein-protein interactions in pure mammalian cell lysates.