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Quantitative Analysis of How RET Receptor Activation is Coupled to ERK and Akt Signaling
Author(s) -
Li Simin,
Noorbaksh Javad,
Riera Thomas V.,
Mehta Pankaj,
Whitty Adrian
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1041.1
Subject(s) - protein kinase b , crosstalk , mapk/erk pathway , signal transduction , receptor , glial cell line derived neurotrophic factor , receptor tyrosine kinase , phosphorylation , microbiology and biotechnology , growth factor receptor , kinase , chemistry , tyrosine kinase , biology , cancer research , neurotrophic factors , biochemistry , physics , optics
The receptor tyrosine kinase RET is activated by growth factors of the GDNF family in conjunction with one of four glycosylphosphatidylinositol‐linked co‐receptors, GFRα1–4. Like other growth factor receptors, our quantitative knowledge of RET function, and particularly how RET activation is quantitatively and temporally coupled to downstream signaling events, remains very rudimentary. We have addressed this knowledge gap by using ELISA assays to precisely measure the phosphorylation levels of RET receptor, ERK1/2 and Akt that are observed upon stimulating mouse NB41A3 neuroblastoma cells expressing RET and GFRα3 with various concentrations of the growth factor artemin for different periods of time. We also use small molecule inhibitors of different steps in these signaling pathways to gain additional information by orthogonally perturbing the system, and to probe crosstalk between the ERK and Akt pathways. We extracted key features of the RET, ERK1/2 and Akt activation processes, such as initial activation rate, maximum and steady‐state signal amplitude, and signal duration, and tested the resulting relationships against models reflecting different connection topologies between these events. The results shed important new light on how activation of a growth factor receptor is quantitatively and temporally coupled to key events in divergent downstream signaling pathways. (GM087469)