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    • 5. 发明授权
    • Assay and reagents for detecting inhibitors of ubiquitin-dependent
degradation of cell cycle regulatory proteins
    • 用于检测细胞周期调节蛋白泛素依赖性降解抑制剂的测定和试剂
    • US5726025A
    • 1998-03-10
    • US425299
    • 1995-04-20
    • Marc W. KirschnerRandall W. KingJan-Michael Peters
    • Marc W. KirschnerRandall W. KingJan-Michael Peters
    • C12Q1/25G01N33/68C12Q1/48G01N33/567
    • G01N33/6842C12Q1/25G01N33/68
    • The present invention provides a systematic and practical approach for the identification of candidate agents able to inhibit ubiquitin-mediated degradation of a cell-cycle regulatory protein, such as cyclins. One aspect of the present invention relates to a method for identifying an inhibitor of ubiquitin-mediated proteolysis of a cell-cycle regulatory protein by (i) providing a ubiquitin-conjugating system that includes the regulatory protein and ubiquitin under conditions which promote the ubiquitination of the target protein, and (ii) measuring the level of ubiquitination of the subject protein brought about by the system in the presence and absence of a candidate agent. A decrease in the level of ubiquitin conjugation is indicative of an inhibitory activity for the candidate agent. The level of ubiquitination of the regulatory protein can be measured by determining the actual concentration of protein:ubiquitin conjugates formed; or inferred by detecting some other quality of the subject protein affected by ubiquitination, including the proteolytic degradation of the protein.
    • 本发明提供了一种用于鉴定能够抑制泛素介导的细胞周期调节蛋白如细胞周期蛋白降解的候选试剂的系统和实用的方法。 本发明的一个方面涉及通过(i)在促进泛素化的条件下提供包含调节蛋白和泛素的泛素缀合系统来鉴定泛素介导的细胞周期调节蛋白的蛋白水解抑制剂的方法 目标蛋白质,以及(ii)测量在存在和不存在候选药剂的情况下由系统产生的受试蛋白质的泛素化水平。 泛素缀合水平的降低表明候选试剂的抑制活性。 调节蛋白的泛素化水平可以通过测定蛋白质的实际浓度来测量:泛素缀合物的形成; 或通过检测受泛素化影响的受试蛋白的一些其他质量,包括蛋白质的蛋白水解降解来推断。