title: Polyamines stimulate the phosphorylation of phosphatidylinositol in membranes from A431 cells.
abstract: The phosphorylation of phosphatidylinositol in plasma membranes from A431 cells was investigated using [gamma-32P]ATP as the substrate. Phosphatidylinositol 4-phosphate was found to be the major product after an incubation time of 5-10 min. Little, if any, phosphatidylinositol 4,5-bisphosphate was found under these conditions. Epidermal growth factor (EGF) had no effect on the formation of phosphatidylinositol 4-phosphate or phosphatidylinositol 4,5-bisphosphate. On the other hand, the polyamines spermidine and spermine stimulated the phosphatidylinositol kinase activity about eightfold yielding almost exclusively phosphatidylinositol 4-phosphate as the reaction product. Half-maximum stimulation by spermidine occurred under near physiological conditions (1.5 mM). Furthermore various proteins and amino acid polymers containing clustered basic amino acid residues (e.g. histones and polylysine) stimulated the formation of phosphatidylinositol 4-phosphate to a similar extent. Half-maximal concentrations for the activation were considerably lower ranging from 1.5 microM to 80 microM. The ATP specificity of the phosphatidylinositol kinase(s) was investigated with a small set of selected ATP derivatives. In the presence of spermidine the specificity changed significantly indicating that (a) spermidine acts on a kinase and not on a phosphatase, (b) this activity is distinct from the EGF-receptor protein kinase activity. The results do not suggest an involvement of the EGF receptor in the growth-factor-dependent formation of phosphatidylinositol phosphates. It is proposed that the phosphorylation of phosphatidylinositol by polyamines might be a mechanism to replenish the pool of inositolphospholipids.
mesh_terms: D019870:1-Phosphatidylinositol 4-Kinase; D000255:Adenosine Triphosphate / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols / Q000096:biosynthesis*; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism; D011073:Polyamines / Q000494:pharmacology*; D013379:Substrate Specificity
chemical_list: D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; D011073:Polyamines; C037178:phosphatidylinositol 4-phosphate; D004815:Epidermal Growth Factor; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D019870:1-Phosphatidylinositol 4-Kinase