Chapter title |
Identification of Receptor Tyrosine Kinase Inhibitors Using Cell Surface Biotinylation and Affinity Isolation
|
---|---|
Chapter number | 8 |
Book title |
VEGF Signaling
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2917-7_8 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2916-0, 978-1-4939-2917-7
|
Authors |
Antony M. Latham, Jayakanth Kankanala, Colin W. G. Fishwick, Sreenivasan Ponnambalam, Latham, Antony M., Kankanala, Jayakanth, Fishwick, Colin W. G., Ponnambalam, Sreenivasan |
Abstract |
The mammalian vascular endothelial growth factor receptor tyrosine kinases (VEGFRs) bind circulating growth factors and regulate the process of angiogenesis. The discovery of new small molecules that target the enzymatic activity of the VEGFR family as potential antiangiogenic drugs is of much commercial interest in the pharmaceutical sector. Here, we describe the use of a combined cell surface biotinylation and affinity isolation procedure to monitor ligand-stimulated VEGFR trafficking in endothelial cells, in which novel VEGFR inhibitors from chemical libraries can be identified by their ability to inhibit receptor internalization. Unlike a traditional cell-free enzyme activity assay, such a cell-based approach provides a physiologically relevant readout of inhibitor activity. In this example, we use the VEGF-A-VEGFR-2 axis and the well-characterized tyrosine kinase inhibitor sunitinib as a working model; however this technique is highly applicable for the identification of inhibitors to other receptor tyrosine kinases. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 8 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Professor | 2 | 25% |
Professor > Associate Professor | 2 | 25% |
Student > Ph. D. Student | 1 | 13% |
Researcher | 1 | 13% |
Unknown | 2 | 25% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 3 | 38% |
Agricultural and Biological Sciences | 1 | 13% |
Medicine and Dentistry | 1 | 13% |
Unknown | 3 | 38% |