A high-content assay to screen for modulators of EGFR function Journal Article
| Authors: | Antczak, C.; Djaballah, H. |
| Article Title: | A high-content assay to screen for modulators of EGFR function |
| Abstract: | Cell-based assays have the potential and advantage to identify cell-permeable modulators of kinase function, and hence provide an alternative to the conventional enzymatic activity-driven discovery approaches that rely on purifi ed recombinant kinase catalytic domains. Here, we describe a domain-based high- content biosensor approach to study endogenous EGFR activity whereby EGF-induced receptor activation, subsequent traffi cking, and internalization are imaged and quantifi ed using time-dependent granule formation in cells. This method can readily be used to search for EGFR modulators in both chemical and RNAi screening; with potential applicability to other receptor tyrosine kinases. © Springer Science+Business Media New York 2016. |
| Keywords: | controlled study; genetics; microscopy; protein conformation; metabolism; proto oncogene; protein kinase inhibitor; epidermal growth factor receptor; neoplasm proteins; green fluorescent protein; small interfering rna; protein binding; rna, small interfering; rna interference; receptor, epidermal growth factor; enzyme activation; drug discovery; drug development; molecular library; small molecule libraries; high-content assay; cell line, tumor; inhibitor; protein tyrosine kinase; phosphorylation; assay; protein kinase inhibitors; protein processing; protein processing, post-translational; screening; tumor protein; tumor cell line; protein transport; green fluorescent proteins; fluorescence microscopy; microscopy, fluorescence; receptor protein-tyrosine kinases; egfr; image processing, computer-assisted; image processing; genetic procedures; isolation and purification; ultrastructure; internalization; egfr protein, human; grb2 adaptor protein; growth factor receptor bound protein 2; indicators and reagents; kinase; rnai; sirna; genes, erbb-1; procedures; hts; biosensing techniques; cell granule; dyes, reagents, indicators, markers and buffers; drug effects; cytoplasmic granules; biosensor; iressa; cancer; humans; human; priority journal; article; inca2000; grb2 protein, human; src homology domain; src homology domains; antagonists and inhibitors; inca3000; high content assay |
| Journal Title: | Methods in Molecular Biology |
| Volume: | 1360 |
| ISSN: | 1064-3745 |
| Publisher: | Humana Press Inc |
| Date Published: | 2016-01-01 |
| Start Page: | 97 |
| End Page: | 106 |
| Language: | English |
| DOI: | 10.1007/978-1-4939-3073-9_8 |
| PUBMED: | 26501905 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Chapter 8 in "Kinase Screening and Profiling" (ISBN: 978-1-4939-3072-2) -- Article -- Export Date: 23 August 2016 -- Source: Scopus |
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