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@ARTICLE{Mohanakumar:911640,
author = {Mohanakumar, Shilpa and Lee, Namkyu and Wiegand, Simone},
title = {{C}omplementary {E}xperimental {M}ethods to {O}btain
{T}hermodynamic {P}arameters of {P}rotein {L}igand
{S}ystems},
journal = {International journal of molecular sciences},
volume = {23},
number = {22},
issn = {1422-0067},
address = {Basel},
publisher = {Molecular Diversity Preservation International},
reportid = {FZJ-2022-04896},
pages = {14198 -},
year = {2022},
abstract = {In recent years, thermophoresis has emerged as a promising
tool for quantifying biomolecular interactions. The
underlying microscopic physical effect is still not
understood, but often attributed to changes in the hydration
layer once the binding occurs. To gain deeper insight, we
investigate whether non-equilibrium coefficients can be
related to equilibrium properties. Therefore, we compare
thermophoretic data measured by thermal diffusion forced
Rayleigh scattering (TDFRS) (which is a non-equilibrium
process) with thermodynamic data obtained by isothermal
titration calorimetry (ITC) (which is an equilibrium
process). As a reference system, we studied the chelation
reaction between ethylenediaminetetraacetic acid (EDTA) and
calcium chloride (CaCl2) to relate the thermophoretic
behavior quantified by the Soret coefficient ST to the
Gibb’s free energy ΔG determined in the ITC experiment
using an expression proposed by Eastman. Finally, we have
studied the binding of the protein Bovine Carbonic Anhydrase
I (BCA I) to two different benzenesulfonamide derivatives:
4-fluorobenzenesulfonamide (4FBS) and
pentafluorobenzenesulfonamide (PFBS). For all three systems,
we find that the Gibb’s free energies calculated from ST
agree with ΔG from the ITC experiment. In addition, we also
investigate the influence of fluorescent labeling, which
allows measurements in a thermophoretic microfluidic cell.
Re-examination of the fluorescently labeled system using ITC
showed a strong influence of the dye on the binding
behavior.},
cin = {IBI-4},
ddc = {540},
cid = {I:(DE-Juel1)IBI-4-20200312},
pnm = {5241 - Molecular Information Processing in Cellular Systems
(POF4-524)},
pid = {G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)16},
pubmed = {36430678},
UT = {WOS:000887546500001},
doi = {10.3390/ijms232214198},
url = {https://juser.fz-juelich.de/record/911640},
}