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Using affinity capillary electrophoresis and computational models for binding studies of heparinoids with p‐selectin and other proteins
Author(s) -
Mozafari Mona,
Balasupramaniam Shantheya,
Preu Lutz,
El Deeb Sami,
Reiter Christian G.,
Wätzig Hermann
Publication year - 2017
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201600480
Subject(s) - capillary electrophoresis , affinity electrophoresis , chemistry , chromatography , selectin , electrophoresis , biophysics , biochemistry , affinity chromatography , biology , enzyme , organic chemistry , adhesion
A fast and precise affinity capillary electrophoresis (ACE) method has been developed and applied for the investigation of the binding interactions between P‐selectin and heparinoids as potential P‐selectin inhibitors in the presence and absence of calcium ions. Furthermore, model proteins and vitronectin were used to appraise the binding behavior of P‐selectin. The normalized mobility ratios (∆R/R f ), which provided information about the binding strength and the overall charge of the protein–ligand complex, were used to evaluate the binding affinities. It was found that P‐selectin interacts more strongly with heparinoids in the presence of calcium ions. P‐selectin was affected by heparinoids at the concentration of 3 mg/L. In addition, the results of the ACE experiments showed that among other investigated proteins, albumins and vitronectin exhibited strong interactions with heparinoids. Especially with P‐selectin and vitronectin, the interaction may additionally induce conformational changes. Subsequently, computational models were applied to interpret the ACE experiments. Docking experiments explained that the binding of heparinoids on P‐selectin is promoted by calcium ions. These docking models proved to be particularly well suited to investigate the interaction of charged compounds, and are therefore complementary to ACE experiments.