文章摘要
张 慧,黄 卉,李 娜,杜培革,石太平.针对药靶基因PDCD10和MST4抑制剂体外筛选方法的建立及化合物的筛选[J].南京医科大学学报,2010,(11):1527~1532
针对药靶基因PDCD10和MST4抑制剂体外筛选方法的建立及化合物的筛选
The establishment of drug targets PDCD10 and MST4 inhibitors screening method in vitro and the compounds screening
投稿时间:2010-04-21  
DOI:10.7655
中文关键词: PDCD10  MST4  抑制剂  ELISA  Elk1信号通路
英文关键词: PDCD10  MST4  inhibitors  ELISA  Elk1 signaling pathway
基金项目:国家高技术研究发展计划(863)“十一五”重大专项资助课题(2006AA02A305),重点项目资助课题(2006AA020501),“重点新药创制”科技重大专项资助课题(2009ZX09503-004)
作者单位
张 慧 北华大学药学院,吉林 吉林 132013 
黄 卉 国家人类基因组北方研究中心,北京 100176 
李 娜 国家人类基因组北方研究中心,北京 100176 
杜培革 北华大学药学院,吉林 吉林 132013 
石太平 国家人类基因组北方研究中心,北京 100176 
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中文摘要:
      目的:以人程序性细胞死亡分子10(homo sapiens programmed cell death 10, PDCD10)和Ste20激酶家族成员MST4(Mst3 and SOK1-related kinase)为靶标,建立体外小分子抑制剂筛选方法?方法:将PDCD10和MST4的开放读码框插入到原核表达载体pGEX4T-2,采用亲和层析纯化法纯化重组人GST-PDCD10和GST-MST4蛋白并通过Western blot进行鉴定?利用ELISA方法对重组蛋白进行活性检测?利用该ELISA方法对825个小分子化合物进行筛选,并通过其对Elk1通路的作用进行验证?结果:成功建立了针对药靶基因PDCD10和MST4抑制剂的体外筛选方法,筛选得到1个对PDCD10和MST4活性有抑制作用的化合物?结论:该方法具有较好的平行性和可重复性?筛选得到的化合物可以抑制PDCD10和MST4对Elk1通路的活化作用?
英文摘要:
      Objective:Taking PDCD10 (homo sapiens programmed cell death 10) and MST4 (Mst3 and SOK1-related kinase) as drug targets, to establish a low molecular weight inhibitors screening method in vitro. Methods:The open reading frames of PDCD10 and MST4 were reconstructed into the prokaryotic expressive vector pGEX4T-2. The recombinant proteins were purified using GST affinity chromatograph, and then analyzed using Western blot. ELISA assay was utilized to detect the activity of purified proteins of GST-PDCD10 and GST-MST4 and screen for the inhibitor in 825 compounds, which were verified by the dual luciferase reporter gene assay in Elk1 signaling pathway. Results:Drug targets PDCD10 and MST4 inhibitors screening method was established successfully in vitro. One compound screened by this method was confirmed to inhibit the activity of PDCD10 and MST4. Conclusion: The method is of better parallelism and stability. The screened compound could also inhibit the activity of PDCD10 and MST4 in Elk1 signaling pathway.
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