Development of a RapidFire mass spectrometry assay and a fluorescence assay for the discovery of kynurenine aminotransferase II inhibitors to treat central nervous system disorders
| Autor: | Dianlin Xie, Litao Zhang, Basanth Laksmaiah, Prasad Krishnamurthy, Mark R. Witmer, Marilyn Paul, Kaushik Ghosh, Sumit Gupta, Lisa M. Kopcho, Michael F. Parker, Jeffrey Tredup, Lynn M. Abell, Hao Lu |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Central nervous system Biophysics Drug Evaluation Preclinical Mass spectrometry Kynurenic Acid Biochemistry Mass Spectrometry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Kynurenic acid Central Nervous System Diseases High-Throughput Screening Assays medicine Humans Enzyme Inhibitors Molecular Biology Transaminases Enzyme Assays chemistry.chemical_classification biology Brain Kynurenine aminotransferase II Cell Biology Enzyme assay 030104 developmental biology medicine.anatomical_structure Enzyme Spectrometry Fluorescence chemistry biology.protein biology.gene 030217 neurology & neurosurgery Kynurenine |
| Zdroj: | Analytical biochemistry. 501 |
| ISSN: | 1096-0309 |
| Popis: | Kynurenine aminotransferases convert kynurenine to kynurenic acid and play an important role in the tryptophan degradation pathway. Kynurenic acid levels in brain have been hypothesized to be linked to a number of central nervous system (CNS) disorders. Kynurenine aminotransferase II (KATII) has proven to be a key modulator of kynurenic acid levels in brain and, thus, is an attractive target to treat CNS diseases. A sensitive, high-throughput, label-free RapidFire mass spectrometry assay has been developed for human KATII. Unlike other assays, this method is directly applicable to KATII enzymes from different animal species, which allows us to select proper animal model(s) to evaluate human KATII inhibitors. We also established a coupled fluorescence assay for human KATII. The short assay time and kinetic capability of the fluorescence assay provide a useful tool for orthogonal inhibitor validation and mechanistic studies. |
| Databáze: | OpenAIRE |
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