Eglen Richard M, Singh Rajendra
Research and Development, DiscoveRx Corp., 42501 Albrae St., Suite 100, Fremont, CA 94538, USA.
Comb Chem High Throughput Screen. 2003 Jun;6(4):381-7. doi: 10.2174/138620703106298473.
In this review, the applications of beta galactosidase complementation are described. alpha Complementation is a naturally occurring process in bacteria and in engineered cells, and can also occur in eukaryotic cells. Two forms of alpha complementation have been used in high throughput screening (HTS), in which interacting fragments complement with either low or high affinity. Low affinity complementation is used to monitor protein protein interactions, such as those occurring in homodimerization of the epidermal growth factor receptor (EGFR), and provides a robust screen for detection of EGFR inhibitors. High affinity complementation provides the basis for several HTS assays, in which analytes, such as cAMP or IP(3), are detected in crude cell lysates. A development of the latter approach is protein labeling, providing for measurement of cell protein expression and trafficking. Collectively, the use of beta galactosidase complementation provides a novel and flexible technology for highly sensitive, homogeneous HTS assay development.
在本综述中，描述了β-半乳糖苷酶互补的应用。α-互补是细菌和工程细胞中自然发生的过程，也可发生在真核细胞中。两种形式的α-互补已用于高通量筛选（HTS），其中相互作用的片段以低亲和力或高亲和力互补。低亲和力互补用于监测蛋白质-蛋白质相互作用，例如表皮生长因子受体（EGFR）同源二聚化中发生的相互作用，并为检测EGFR抑制剂提供了强大的筛选方法。高亲和力互补为几种HTS分析提供了基础，其中可在粗细胞裂解物中检测诸如环磷酸腺苷（cAMP）或肌醇三磷酸（IP₃）等分析物。后一种方法的一个发展是蛋白质标记，可用于测量细胞蛋白质表达和运输。总的来说，β-半乳糖苷酶互补的使用为高灵敏度、均相HTS分析方法的开发提供了一种新颖且灵活的技术。
