A microfluidics-based mobility shift assay to discover new tyrosine phosphatase inhibitors
| Autor: | Alexander Scheer, Christèle Frémaux, Wolfgang H. B. Sauer, Dominique Perrin, Dominique Besson |
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| Rok vydání: | 2006 |
| Předmět: |
0301 basic medicine
Microfluidics Drug Evaluation Preclinical Peptide Electrophoretic Mobility Shift Assay Protein tyrosine phosphatase Biology 01 natural sciences Biochemistry Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Structure-Activity Relationship Structure–activity relationship Electrophoretic mobility shift assay chemistry.chemical_classification Phosphopeptide Tyrosine phosphorylation 0104 chemical sciences 010404 medicinal & biomolecular chemistry 030104 developmental biology Drug development chemistry Peptidyl Transferases Molecular Medicine Protein Tyrosine Phosphatases Vanadates Biotechnology |
| Zdroj: | Journal of biomolecular screening. 11(8) |
| ISSN: | 1087-0571 |
| Popis: | Protein tyrosine phosphatases (PTPs) play key roles in regulating tyrosine phosphorylation levels in cells. Since the discovery of PTP1B as a major drug target for diabetes and obesity, PTPs have emerged as a new and promising class of signaling targets for drug development in a variety of therapeutic areas. The routine use of generic substrate 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP) in our hands led to the discovery of very similar and often not very selective molecules. Therefore, to increase the chances to discover novel chemical scaffolds, a side-by-side comparison between the DiFMUP assay and a chip-based mobility shift assay with a specific phosphopeptide was performed, on 1 PTP, using a focused set of compounds. Assay robustness and sensitivity were comparable for both the DiFMUP and mobility shift assays. The off-chip mobility shift assay required a longer development time because of identification, synthesis, and characterization of a specific peptide, and its cost per point was higher. However, although most potent scaffolds found with the DiFMUP assay were confirmed in the mobility shift format, the off-chip mobility shift assay led to the identification of previously unidentified chemical scaffolds with improved druglike properties. |
| Databáze: | OpenAIRE |
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