金黄色葡萄球菌色氨酰-tRNA合成酶抑制剂体外筛选模型的建立
Establishment of in vitro models for screening Staphylococcal aureus tryptophanyl-tRNA synthetase inhibitors
目的 建立金黄色葡萄球菌色氨酰-tRNA合成酶抑制剂的体外筛选模型和方法。方法 建立检测底物ATP减少量的ATP-luminescence法和检测产物焦磷酸生成的PPi-Absorbance法，并对方法进行验证。结果 对两种方法优化后的底物浓度分别为8 μM ATP和40 μM L-色氨酸，酶最优浓度为2 μg；验证酶活检测方法建立成功 。结论 所建立的金黄色葡萄球菌色氨酰-tRNA合成酶抑制剂的体外筛选模型可以为快速-高通量筛选抑制剂提供技术平台。
Objective: Establishing in vitro models and methods for screening inhibitors of Staphylococcal aureus tryptophanyl-tRNA synthetase. Methods: Determine the amount of remaining substrate ATP by detecting luminescence and determine the product diphosphate by monitoring absorbance at 340 nm. Results: For tryptophanyl-tRNA synthetase calalyzed reaction, the optimized concentrations of ATP, L-tryptophan and SaWRS are 8 μM, 40 μM and 2 μg, respectively. The IC50 of compound W-23 against SaWRS is 9.7-10.5 μM. Conclusion: The in vitro models established in the study can provide a fast and high-throughput platform for screening Staphylococcal aureus tryptophanyl-tRNA synthetase inhibitors.
瞿涤、吴旸
基础医学生物科学研究方法、生物科学研究技术微生物学
病原生物学葡萄球菌色氨酰-tRNA合成酶抑制剂体外筛选
Medical MicrobiologyStaphylococcustryptophanyl-tRNA synthetaseinhibitor screening
瞿涤,吴旸.金黄色葡萄球菌色氨酰-tRNA合成酶抑制剂体外筛选模型的建立[EB/OL].(2014-02-26)[2025-05-23].http://www.paper.edu.cn/releasepaper/content/201402-515.点此复制
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