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Inhibition of Epidermal Growth Factor stimulated ERK phosphorylation by inositol trisphosphate kinase inhibitor
Author(s) -
Sekar M. Chandra,
Leloup Ludovic,
Shao Hanshuang,
Wells Alan
Publication year - 2009
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.23.1_supplement.889.1
Subject(s) - mapk/erk pathway , epidermal growth factor , kinase , microbiology and biotechnology , phosphorylation , inositol , chemistry , biology , biochemistry , receptor
We have used an IP 3 ‐kinase [IP3K] inhibitor, 2‐trifluoromethyl [6‐(4‐nitrobenzyl)‐purine] to dissect the role of inositol (1,3,4,5)P 4 [IP 4 ] in epidermal growth factor [EGF] signaling. Screening of cDNA with IP 3 kinase enzyme primers indicated that at least one subtype of IP3K is present in both endothelial [EC] and fibroblast [FB] cells. Quiescent EC and FB cells were stimulated with EGF in presence and absence of IP3K inhibitor. Cell were lysed, and proteins separated by SDS gel electrophoresis; the western blots were probed with polyclonal antibody. The phosphorylation level of ERK stimulated by EGF for short time period (15 min) was significantly inhibited by 10 µM IP3K inhibitor. Phospho‐ERK was blocked by 36% in EC and 28% in FB. No consistent inhibition was observed following long term [16 h] EGF stimulation. As ERK‐phosphorylation is essential for m‐calpain activation, rear cell detachment and cell motility, our data suggests that IP 4 may play a role in modulating one or more of these steps. (Supported by a grant from the National Institute of General Medical Sciences (NIH) to Alan Wells).