Evidence for a rapid stimulation of tyrosine kinase activity by prolactin in Nb2 rat lymphoma cells.
Studies were designed to determine if the activation of tyrosine kinases may be involved in the signal transduction pathway for PRL. Tyrosyl phosphorylation of cellular proteins was evaluated by western blot analysis of Nb2 cell proteins employing an antibody to phosphotyrosine. Physiological concentrations of ovine PRL (oPRL) had a pronounced effect on the tyrosyl phosphorylation of a 121 kDa protein. Increased tyrosyl phosphorylation of the 121 kDa protein was detectable with concentrations of oPRL as low as 0.5 ng/ml. Consistent with oPRL acting through a PRL receptor, hGH also stimulated tyrosyl phosphorylation of the 121 kDa protein when tested at concentrations between 5 and 20 ng/ml. In time course experiments, increased tyrosyl phosphorylation of the 121 kDa protein was apparent after a 5 min incubation with 20 ng/ml hGH, and maintained for at least one h. At higher concentrations of hGH (200 ng/ml), increased phosphorylation of the 121 kDa protein was clearly evident after only 1 min, indicating that tyrosyl phosphorylation of cellular proteins is an early event following ligand binding to the PRL receptor. Increased tyrosyl phosphorylation of proteins of 40, 90 and 55-65 kDa was also evident after incubation with hGH for 10, 10, and 60 min respectively. These findings are consistent with PRL-dependent tyrosine kinase activation being an early and perhaps initiating event in the signal transduction pathway for PRL in Nb2 cells.
Rillema
J A
JA
Department of Physiology, Wayne State University School of Medicine, Detroit, MI 48201.
Campbell
G S
GS
Lawson
D M
DM
Carter-Su
C
C
D000818:Animals; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008223:Lymphoma / Q000201:enzymology*; D008970:Molecular Weight; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011388:Prolactin / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats; D011981:Receptors, Prolactin / Q000378:metabolism; D011986:Receptors, Somatotropin / Q000378:metabolism; D014407:Tumor Cells, Cultured
D010750:Phosphoproteins; D011981:Receptors, Prolactin; D011986:Receptors, Somatotropin; D011388:Prolactin; D011505:Protein-Tyrosine Kinases