Development and comprehensive comparison of two on-line capillary electrophoretic methods for beta-secretase inhibitor screening
| Authors | |
|---|---|
| Year of publication | 2017 |
| Type | Article in Periodical |
| Magazine / Source | Journal of Chromatography A |
| MU Faculty or unit | |
| Citation | |
| web | https://doi.org/10.1016/j.chroma.2017.08.065 |
| Doi | http://dx.doi.org/10.1016/j.chroma.2017.08.065 |
| Field | Biochemistry |
| Keywords | capillary electrophoresis; Alzheimer's disease; beta-secretase; kinetics; inhibition |
| Description | Alzheimer’s disease is the most common cause of dementia, afflicting over 34 million patients worldwide. Since beta-secretase is a rate-limiting enzyme of the production of neurotoxic beta-amyloid peptide oligomers abnormally accumulated in the affected brain tissue, its specific inhibition appears to be a promising approach to slowing down or even stopping the progression of the disease. Hence two on-line capillary electrophoretic methods for studies of beta-secretase activity based on the principles of transverse diffusion of laminar flow profiles and electrophoretically mediated microanalysis were developed, both using a simple unlabeled peptide substrate and UV detection. The optimized procedures were thoroughly validated and applied for determining the enzyme’s kinetic parameters and the inhibition characteristics of two potent probe inhibitors. The resulting values were found to be comparable to literature data obtained with other analytical techniques. The suitability of the employed methodologies for different experimental designs is discussed on the basis of a statistical evaluation of the experimental data. The presented methods constitute a miniaturized and fully automated tool, which should be suitable for kinetic and inhibition studies of beta-secretase as a target for Alzheimer’s disease drug discovery in the early stages of the development of a new drug. |
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