Microscale Thermophoresis: Interactions of Proteins Small Molecules Nucleic Acids and Vesicles

摘要

This work gives an overview on a new Technology for the measurement of biomolecule interaction that is termed Microscale Thermophoresis (MST). The term Microscale Thermophoresis refers to the directed movement of molecules in optically generated microscopic temperature gradients. This thermophoretic movement is determined by the entropy of the hydration shell around the molecules. Almost all interactions between molecules and virtually any biochemical process related to a change in size, stability and conformation of molecules alters this hydration shell and can be quantified. Such changes allow quantification of binding affinities of proteins, nucleic acids and small molecules as well as measurement of enzymatic activities with MST. In addition also functional studies of small molecule inhibitors are possible. The microscopic temperature gradient is generated by an IR-Laser, which is strongly absorbed by water. The readout method of the interaction analysis is based on fluorescence: intrinsic fluorescence of proteins can be used as well as proteins expressed with GFP/YFP/RFP and also dye labeled biomolecules. In this presentation we will describe the technical details and the benefits of the Microscale Thermophoresis technology platform. We will show examples for interaction measurements ranging from protein -ribosome, protein -protein, small molecule –receptor binding to studies where the interactions between receptor containing vesicles and proteins are analyzed.