Wittmann, Hans-Joachim and Strasser, Andrea (2017) Competitive association binding kinetic assays: a new tool to detect two different binding orientations of a ligand to its target protein under distinct conditions? NAUNYN-SCHMIEDEBERGS ARCHIVES OF PHARMACOLOGY, 390 (6). pp. 595-612. ISSN 0028-1298, 1432-1912
Full text not available from this repository. (Request a copy)Abstract
Within the last years, for several ligands, binding to G protein-coupled receptors or other target proteins, a binding of the ligand in two different orientations is described. One appropriate experimental technique to detect two different binding orientations is the crystallization of the ligand-protein-complex, but crystallization and subsequent X-ray analysis do not belong to the routine methods. By traditional competitive radioligand equilibrium binding assays, it is not possible to detect or to distinguish between two different binding orientations, but there is a possibility to identify two different binding orientations by performing kinetic competitive radioligand-binding assays. To study the limitations of this new technique, the related differential equations were defined and solved numerically for 8 different sets of rate constants, also considering an experimental error up to similar to 10%. In principal, the kinetic competitive radioligand binding assay is a suitable technique to detect two different ligand binding orientations. However, the present study shows that this is only possible under distinct conditions: (1) the rate constants of dissociation for both binding orientations of the cold ligand should at least be >> 10-fold different to each other and (2) the experimental error should be as small as possible. Although there are some limitations for the experimental usability of this method, it is worthwhile to perform kinetic competitive binding assays, especially if there are hints for two binding orientations of a ligand, e.g. based on molecular modelling studies.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | RESIDENCE TIME; DRUG TARGETS; RECEPTOR; HISTAPRODIFENS; ACTIVATION; MECHANISM; AGONISTS; OPINION; Competitive binding kinetics; Different binding orientations; Rate constant of association; Rate constant of dissociation |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institute of Pharmacy |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Dec 2018 13:10 |
| Last Modified: | 18 Feb 2019 13:12 |
| URI: | https://pred.uni-regensburg.de/id/eprint/849 |
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