Beta galactosidase enzyme fragment complementation as a novel technology for high throughput screening.
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.
Eglen
Richard M
RM
Research and Development, DiscoveRx Corp., 42501 Albrae St., Suite 100, Fremont, CA 94538, USA. reglen@discoverx.com
Singh
Rajendra
R
D000818:Animals; D002460:Cell Line; D000242:Cyclic AMP / Q000032:analysis / Q000378:metabolism; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology; D005816:Genetic Complementation Test; D015544:Inositol 1,4,5-Trisphosphate / Q000032:analysis / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics; D025941:Protein Interaction Mapping / Q000379:methods; D011506:Proteins / Q000378:metabolism; D001616:beta-Galactosidase / Q000737:chemistry* / Q000235:genetics*
D010446:Peptide Fragments; D011506:Proteins; D015544:Inositol 1,4,5-Trisphosphate; D000242:Cyclic AMP; D001616:beta-Galactosidase