TY - JOUR
T1 - Using two-site binding models to analyze microscale thermophoresis data
AU - Tso, Shih Chia
AU - Chen, Qiuyan
AU - Vishnivetskiy, Sergey A.
AU - Gurevich, Vsevolod V.
AU - Iverson, T. M.
AU - Brautigam, Chad A
N1 - Funding Information:
Part of the research in this report was supported by grants to VVG ( GM077561 , GM109955 , (these two R01 grants were replaced by R35 GM122491 ), and EY011500 ) and TMI ( GM120569 , GM061606 , AI106987 , and DA043680 ) from the National Institutes of Health , a Vanderbilt Discovery Grant (TMI/VVG), and the Vanderbilt Core Grant in Vision Research ( P30EY008126 ).
Publisher Copyright:
© 2017 Elsevier Inc.
PY - 2018/1/1
Y1 - 2018/1/1
N2 - The emergence of microscale thermophoresis (MST) as a technique for determining the dissociation constants for bimolecular interactions has enabled these quantities to be measured in systems that were previously difficult or impracticable. However, most models for analyses of these data featured the assumption of a simple 1:1 binding interaction. The only model widely used for multiple binding sites was the Hill equation. Here, we describe two new MST analytic models that assume a 1:2 binding scheme: the first features two microscopic binding constants (KD(1) and KD(2)), while the other assumes symmetry in the bivalent molecule, culminating in a model with a single macroscopic dissociation constant (KD,M) and a single factor (α) that accounts for apparent cooperativity in the binding. We also discuss the general applicability of the Hill equation for MST data. The performances of the algorithms on both real and simulated data are assessed, and implementation of the algorithms in the MST analysis program PALMIST is discussed.
AB - The emergence of microscale thermophoresis (MST) as a technique for determining the dissociation constants for bimolecular interactions has enabled these quantities to be measured in systems that were previously difficult or impracticable. However, most models for analyses of these data featured the assumption of a simple 1:1 binding interaction. The only model widely used for multiple binding sites was the Hill equation. Here, we describe two new MST analytic models that assume a 1:2 binding scheme: the first features two microscopic binding constants (KD(1) and KD(2)), while the other assumes symmetry in the bivalent molecule, culminating in a model with a single macroscopic dissociation constant (KD,M) and a single factor (α) that accounts for apparent cooperativity in the binding. We also discuss the general applicability of the Hill equation for MST data. The performances of the algorithms on both real and simulated data are assessed, and implementation of the algorithms in the MST analysis program PALMIST is discussed.
KW - Arrestin-3
KW - DNA aptamer
KW - Microscale thermophoresis
KW - Protein-ligand interactions
KW - Protein-protein interactions
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U2 - 10.1016/j.ab.2017.10.013
DO - 10.1016/j.ab.2017.10.013
M3 - Article
C2 - 29054528
AN - SCOPUS:85034821788
SN - 0003-2697
VL - 540-541
SP - 64
EP - 75
JO - Analytical Biochemistry
JF - Analytical Biochemistry
ER -