Nerve growth factor stimulates protein tyrosine phosphorylation in PC-12 pheochromocytoma cells.
The cellular actions of nerve growth factor (NGF) and epidermal growth factor (EGF) may be mediated by changes in protein phosphorylation. The tyrosine phosphorylation of two predominant proteins of molecular mass 40 and 42 kDa is seen in PC-12 cells treated with NGF or EGF, correlating with activation of a previously identified serine/threonine protein kinase that phosphorylates microtubule-associated protein (MAP). Stimulation of phosphoprotein (pp) 40 and 42 phosphorylation and MAP kinase activity by NGF but not EGF is selectively attenuated by staurosporine and K-252A. Moreover, the time courses of pp40/42 phosphorylation and MAP kinase activation produced by NGF or EGF are identical. Chromatography of lysates from growth factor-treated cells on ion-exchange or hydrophobic-interaction HPLC resolves MAP kinase into two peaks, neither of which precisely coelutes with pp40 or pp42. One of these peaks (II) exhibits no detectable phosphotyrosine. The other peak (I) has some overlap with pp40. However, the activity residing in both peaks is almost completely inhibited after treatment with alkaline phosphatase, suggesting that, at least, serine/threonine phosphorylation is required for the activity of these enzymes. These data indicate that while tyrosine phosphorylation appears to be a critical early event in NGF action, the role of this modification in activation of MAP kinases remains unclear.
Miyasaka
T
T
Laboratory of Molecular Oncology, Rockefeller University, New York, NY 10021.
Sternberg
D W
DW
Miyasaka
J
J
Sherline
P
P
Saltiel
A R
AR
D000310:Adrenal Gland Neoplasms; D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002460:Cell Line; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009414:Nerve Growth Factors / Q000494:pharmacology*; D010673:Pheochromocytoma; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D014443:Tyrosine
D007328:Insulin; D008869:Microtubule-Associated Proteins; D009414:Nerve Growth Factors; D047428:Protein Kinase Inhibitors; D014443:Tyrosine; D004815:Epidermal Growth Factor; D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases