Molecular recognition via protein–ligand interactions is of fundamental importance to numerous processes in living organisms. Microscale thermophoresis (MST) uses the sensitivity of the thermophoretic response upon ligand binding to access information on the reaction kinetics. Additionally, thermophoresis is promising as a tool to gain information on the hydration layer, as the temperature dependence of the thermodiffusion behaviour is sensitive to solute-solvent interactions. To quantify the influence of structural fluctuations and conformational motion of the protein on the entropy change of its hydration layer upon ligand binding, we combine quasi-elastic incoherent neutron scattering (QENS) and isothermal titration calorimetry (ITC) data from literature. However, preliminary results show that replacing water with deuterated water leads to changes of the thermophoretic measurements, which are similar to the changes observed upon binding by biotin. In order to gain a better understanding of the hydration layer all measurements need to be performed in heavy water. This will open a route to develop a microscopic understanding of the correlation between the strength and number of hydrogen bonds and the thermophoretic behaviour.
Thermodiffusion as a probe of protein hydration for streptavidin and the streptavidin-biotin complex
Doreen Niether, Mona Sarter, Bernd König, Michaela Zamponi, Jörg Fitter, Andreas Stadler, Simone Wiegand; Thermodiffusion as a probe of protein hydration for streptavidin and the streptavidin-biotin complex. AIP Conf. Proc. 22 January 2018; 1929 (1): 020001. https://doi.org/10.1063/1.5021914
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