ANTI-PHOSPHOTYROSINE IMMUNOPRECIPITATION OF PHOSPHATIDYLINOSITOL-3' KINASE-ACTIVITY IN DIFFERENT CELL-TYPES AFTER EXPOSURE TO EPIDERMAL GROWTH-FACTOR

In previous studies from this laboratory, it was shown that mouse epidermal growth factor (mEGF) or insulin increased the labeling of phosphaditylinositol-3,4-bisphosphate (PI-3,4-P2) in MA-10 cells prelabeled with different radioactive precursors (Pignataro, O.P., and Ascoli, M. (1990) J. Biol. Chem. 265, 1718-1723 and Mol. Endocrinol. (1990) 4, 758-765). In order to further characterize this phenomenon we sought to determine if we could use anti-phosphotyrosine antibodies to immunoprecipitate a phosphatidylinositol (PI) kinase activity from MA-10 cells treated with mEGF or insulin. Our data indicate that this is indeed the case, and that the PI kinase precipitated is a PI-3′ kinase. A second cell type, A431 cells, in which we were unable to detect an increase in PI-3,4-P2 labeling when stimulated with mEGF or insulin, was also studied. It was found that, as in MA-10 cells, A431 cells also contain an immunoprecipitable PI-3′ kinase activity that is increased in response to mEGF or insulin. © 1990 Academic Press, Inc.