Analysis of receptor tyrosine kinase internalization using flow cytometry.
The internalization of activated receptor tyrosine kinases (RTKs) by endocytosis and their subsequent down regulation in lysosomes plays a critical role in regulating the duration and intensity of downstream signaling events. Uncoupling of the RTK cMet from ligand-induced degradation was recently shown to correlate with sustained receptor signaling and increased cell tumorigenicity, suggesting that the corruption of these endocytic mechanisms could contribute to increased cMet signaling in metastatic cancers. To understand how cMet signaling for normal cell growth is controlled by endocytosis and how these mechanisms are dysregulated in metastatic cancers, we developed flow cytometry-based assays to examine cMet internalization.
Li
Ning
N
Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, TX, USA.
Hill
Kristen S
KS
Elferink
Lisa A
LA
D001426:Bacterial Proteins / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D006982:Hypertonic Solutions / Q000494:pharmacology; D008024:Ligands; D018613:Microscopy, Confocal; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D013194:Staining and Labeling
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D006982:Hypertonic Solutions; D008024:Ligands; C069761:internalin protein, Bacteria; D019859:Proto-Oncogene Proteins c-met; D020794:Receptor Protein-Tyrosine Kinases