title: Quantification of the effects of mutations on receptor tyrosine kinase (RTK) activation in mammalian cells.
abstract: Single amino acid mutations in receptor tyrosine kinases (RTKs) are known to cause receptor over-activation and disease. Here we present a detailed protocol for the quantification of the effect of mutations on RTK activation in mammalian cells. The activation measurements are based on Western blotting, and involve direct comparison of receptor phosphorylation under conditions that ensure identical expression of wild-type and mutant receptors.
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, 3400 N. Charles Street, Baltimore, MD, 21218, USA.
authors.forename: Lijuan
authors.initials: L
authors.lastname: He
authors.forename: Kalina
authors.initials: K
authors.lastname: Hristova
mesh_terms: D045163:Antibodies, Phospho-Specific / Q000737:chemistry; D015153:Blotting, Western / Q000379:methods*; D004789:Enzyme Activation / Q000187:drug effects; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology*; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D010750:Phosphoproteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D045163:Antibodies, Phospho-Specific; D010750:Phosphoproteins; D016220:Fibroblast Growth Factor 1; C496365:FGFR3 protein, human; D051498:Receptor, Fibroblast Growth Factor, Type 3