Assay Development and Screening for the Identification of Ganglioside GM3 Synthase Inhibitors
| Autor: | Kenji Yamanaka, Yu Takahashi, Yoshiji Hantani, Yuya Azuma |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Sialyltransferase
Biochemistry Chemical library chemistry.chemical_compound Insulin resistance Tandem Mass Spectrometry medicine G(M3) Ganglioside Humans Hypoglycemic Agents Receptor Chromatography High Pressure Liquid Enzyme Assays Ganglioside biology Glycosphingolipid medicine.disease Recombinant Proteins Sialyltransferases High-Throughput Screening Assays Insulin receptor Scintillation proximity assay Diabetes Mellitus Type 2 chemistry biology.protein |
| Zdroj: | Biochemistry. 59:1242-1251 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/acs.biochem.0c00055 |
| Popis: | Ganglioside GM3 is a sialylated membrane-based glycosphingolipid that regulates insulin receptor signaling via direct association with the receptor. The level of expression of GM3 synthase (GM3S) and GM3 is increased in tissues of patients with diabetes and murine models of diabetes, and obesity-induced insulin resistance is attenuated in GM3S-deficient mice. Therefore, GM3S has been considered a therapeutic target for type II diabetes; however, no GM3S inhibitors have been reported to date. In this study, we established a high-throughput scintillation proximity assay that can detect GM3S activity to screen GM3S inhibitors from our original chemical library. We also established methods for detecting the activity of GM3S and another sialyltransferase, ST3Gal3, through direct measurement of the enzyme products using an automatic rapid solid-phase extraction system directly coupled to a mass spectrometer. Consequently, we successfully identified two different chemotypes of GM3S-selective inhibitors with a mixed mode of inhibition. We believe that these compounds can be further developed into drugs to treat or prevent diabetes as well as contribute to the development of the ganglioside research field. |
| Databáze: | OpenAIRE |
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