title: Using affinity capillary electrophoresis and computational models for binding studies of heparinoids with p-selectin and other proteins.
abstract: 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/Rf), 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.
authors.affiliation: Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Braunschweig, Germany.
authors.forename: Mona
authors.initials: M
authors.lastname: Mozafari
authors.affiliation: Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Braunschweig, Germany.
authors.forename: Shantheya
authors.initials: S
authors.lastname: Balasupramaniam
authors.affiliation: Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Braunschweig, Germany.
authors.forename: Lutz
authors.initials: L
authors.lastname: Preu
authors.affiliation: Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Braunschweig, Germany.
authors.forename: Sami
authors.initials: S
authors.lastname: El Deeb
authors.affiliation: bene pharmaChem GmbH & Co.KG, Geretsried, Germany.
authors.forename: Christian G
authors.initials: CG
authors.lastname: Reiter
authors.affiliation: Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Braunschweig, Germany.
authors.forename: Hermann
authors.initials: H
authors.lastname: Wätzig
mesh_terms: D001665:Binding Sites; D002118:Calcium; D003198:Computer Simulation; D019075:Electrophoresis, Capillary; D006496:Heparinoids / Q000737:chemistry*; D007477:Ions; D008024:Ligands; D019007:P-Selectin / Q000737:chemistry*; D011479:Protamines / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*
chemical_list: D006496:Heparinoids; D007477:Ions; D008024:Ligands; D019007:P-Selectin; D011479:Protamines; D011506:Proteins; D002118:Calcium
keywords: MOE docking; Mobility shift affinity capillary electrophoresis; P-Selectin; Protein heparinoid interactions