Screening of 5′-Methylthioadenosine Nucleosidase Enzyme Inhibitors from Traditional Chinese Medicine and Small Molecular Compounds by Capillary Electrophoresis After Enzymatic Reaction at Capillary Inlet
| Autor: | Jie Xu, Ting-Fu Jiang, Kaili Yu, Mei-E Yue |
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| Rok vydání: | 2020 |
| Předmět: |
HEPES
chemistry.chemical_classification Chromatography biology 010405 organic chemistry Capillary inlet 010401 analytical chemistry Organic Chemistry Clinical Biochemistry Phosphate 01 natural sciences Biochemistry Enzyme assay 0104 chemical sciences Analytical Chemistry Electrophoresis chemistry.chemical_compound Capillary electrophoresis Enzyme chemistry Methylthioadenosine nucleosidase biology.protein |
| Zdroj: | Chromatographia. 83:541-548 |
| ISSN: | 1612-1112 0009-5893 |
| DOI: | 10.1007/s10337-020-03873-4 |
| Popis: | A capillary electrophoresis (CE) method was developed for the screening of 5′-methylthioadenosine nucleosidase (MTAN) inhibitors in traditional Chinese drugs and small molecular compounds. In this method, reaction occurred at the capillary inlet during a predetermined waiting period, followed by electrophoretic separation of the reaction substrates and products. Parameters affecting separation efficiency and enzyme activity were evaluated systematically. The optimized conditions were as follows: 30 mM phosphate (pH 3.0) running buffer, 25 kV applied voltage, 20 mM HEPES (pH 6.0) reaction buffer, and 4 min incubation time. Under the optimized conditions, the Michaelis–Menten constant (Km) of MTAN was 3.44 μM, which agreed with the documented data. The proposed method was successfully applied for MTAN inhibitor screening from 21 natural extracts and 8 small molecular compounds. The results demonstrated the practical applicability of the at-inlet reaction-based CE method in screening of MTAN inhibitors from natural products and small molecular compounds. |
| Databáze: | OpenAIRE |
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