Epidermal growth factor alters metabolism of inositol lipids and activity of protein kinase C in mouse embryo palate mesenchyme cells.
Epidermal growth factor (EGF) stimulated mouse embryo palate mesenchyme (MEPM) cells (1) to incorporate [32P]O4(3-) into phosphatidylinositol (PI), phosphatidylcholine, and phosphatidic acid over a period of 60 min; 2) to incorporate [32P]O4(3-) into polyphosphoinositides as a function of time; and 3) to incorporate [32P]O4(-3) into PI, only, as a function of concentration when the period of stimulation was kept short. EGF stimulated the release of radiolabeled inositol phosphates from MEPM cells that had been radiolabeled with [3H]myoinositol. The release of inositol 1-phosphate was sustained over a period of at least 60 min, whereas the release of inositol 1,4-bisphosphate and inositol trisphosphate peaked during the first 10 min of stimulation. EGF also stimulated phosphorylation of an Mr 80,000 protein whose pI, phosphopeptide map, and phosphoamino acid pattern were identical to those of an Mr 80,000 protein phosphorylated in response to phorbol 12-myristate 13-acetate. Mobilization or metabolism of arachidonic acid was not stimulated under the same conditions that permitted EGF to alter inositol lipid metabolism. We interpret these data to mean that 1) in contrast to the findings with some cell lines, alterations in inositol lipid metabolism may be part of the signalling mechanism for EGF in embryonic cells; 2) EGF is capable of activating inositol-dependent signalling pathways leading to activation of protein kinase C in MEPM cells; and 3) mobilization and metabolism of arachidonic acid are not an inherent part of this signalling mechanism.
Chepenik
K P
KP
Department of Anatomy, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Haystead
T A
TA
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D066298:In Vitro Techniques; D007295:Inositol Phosphates / Q000032:analysis; D008648:Mesoderm / Q000378:metabolism; D051379:Mice; D010159:Palate / Q000378:metabolism*; D010449:Peptide Mapping; D010743:Phospholipids / Q000378:metabolism*; D010766:Phosphorylation; D011453:Prostaglandins / Q000032:analysis; D011493:Protein Kinase C / Q000378:metabolism*
D007295:Inositol Phosphates; D010743:Phospholipids; D011453:Prostaglandins; D004815:Epidermal Growth Factor; D011493:Protein Kinase C