Ye Q Z, Nan F J, Hu L Y, Qian Z, Qian J
National Centers for Drug Screening, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Acta Pharmacol Sin. 2000 Nov;21(11):1021-6.
To establish a high-throughput method for inhibitor screening using a recombinant collagenase catalytic domain.
Human collagenase 1 catalytic domain protein was expressed in E coli and used for screening a set of 2720 compounds in a high-throughput fashion.
The screening was accomplished within 2 h and 10 min with consumption of each compound at 4 micrograms. Sixty-six compounds were identified with > 60% inhibitory activity at 20 mg/L, among which 44 compounds were confirmed by subsequent testing at multiple concentrations. The most potent compound showed an IC50 at 4.3 mumol/L, and there were total 15 compounds with IC50 less than 20 mumol/L.
The high-throughput method using the recombinant collagenase is fast, effective and practical in identifying inhibitors.
建立一种使用重组胶原酶催化结构域进行抑制剂筛选的高通量方法。
人胶原酶1催化结构域蛋白在大肠杆菌中表达，并用于高通量筛选一组2720种化合物。
筛选在2小时10分钟内完成，每种化合物的消耗量为4微克。鉴定出66种在20毫克/升时具有>60%抑制活性的化合物，其中44种化合物通过后续多浓度测试得到确认。最有效的化合物的IC50为4.3微摩尔/升，共有15种化合物的IC50小于20微摩尔/升。
使用重组胶原酶的高通量方法在鉴定抑制剂方面快速、有效且实用。
