Studying epigenetic interactions using MicroScale Thermophoresis (MST)

Thomas Schubert; Gernot Längst; Thomas Schubert; Gernot Längst

doi:10.3934/biophy.2015.3.370

AIMS Biophysics

2015, Volume 2, Issue 3: 370-380. doi: 10.3934/biophy.2015.3.370

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Studying epigenetic interactions using MicroScale Thermophoresis (MST)

Thomas Schubert ^{1
,
,},
Gernot Längst ²

1.
2bind GmbH, Josef Engertstraße 13, 93053 Regensburg, Germany;
2.
Biochemie III, Universität Regensburg, Universitätstraße 31, 93053 Regensburg, Germany

Received: 16 July 2015 Accepted: 13 August 2015 Published: 21 August 2015

Abstract
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Epigenetic regulation is based on specific molecular interactions between epigenetic reader, writer and eraser molecules and chromatin. Binding parameters of these interactions such as binding affinities, stoichiometries and thermodynamics are essential for the understanding of the establishment and maintenance of epigenetic networks. The MicroScale Thermophoresis (MST) is a rapid and precise method to characterize epigenetic interactions in solution at microliter scale, requiring low concentrations of the potential interactors. The technology is based on the movement of molecules through temperature gradients, a physical effect referred to as thermophoresis. The thermophoretic movement of a molecule depends on its size, charge and hydration shell. Upon the interaction of two molecules, at least one of these parameters is altered, leading to a change in the movement behavior, which can be used to quantify molecular interactions. MST offers free choice of buffers, also allowing measurements in serum and crude extracts, thereby ensuring optimal reaction conditions. Binding affinities from pM to mM can be measured, perfectly suited to analyze protein/protein, protein/modified peptide and protein/nucleic acid interactions in epigenetics. This review demonstrates the potential of this rapid and versatile technology in the characterization of epigenetic modifiers.
- epigenetics,
- molecular interactions,
- binding affinity,
- MicroScale Thermophoresis (MST)
Citation: Thomas Schubert, Gernot Längst. Studying epigenetic interactions using MicroScale Thermophoresis (MST)[J]. AIMS Biophysics, 2015, 2(3): 370-380. doi: 10.3934/biophy.2015.3.370

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Abstract

Epigenetic regulation is based on specific molecular interactions between epigenetic reader, writer and eraser molecules and chromatin. Binding parameters of these interactions such as binding affinities, stoichiometries and thermodynamics are essential for the understanding of the establishment and maintenance of epigenetic networks. The MicroScale Thermophoresis (MST) is a rapid and precise method to characterize epigenetic interactions in solution at microliter scale, requiring low concentrations of the potential interactors. The technology is based on the movement of molecules through temperature gradients, a physical effect referred to as thermophoresis. The thermophoretic movement of a molecule depends on its size, charge and hydration shell. Upon the interaction of two molecules, at least one of these parameters is altered, leading to a change in the movement behavior, which can be used to quantify molecular interactions. MST offers free choice of buffers, also allowing measurements in serum and crude extracts, thereby ensuring optimal reaction conditions. Binding affinities from pM to mM can be measured, perfectly suited to analyze protein/protein, protein/modified peptide and protein/nucleic acid interactions in epigenetics. This review demonstrates the potential of this rapid and versatile technology in the characterization of epigenetic modifiers.

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