Specificity of phospholipases in methylcholanthrene-transformed mouse fibroblasts activated by bradykinin, thrombin, serum, and ionophore A23187.
Methylcholanthrene-transformed mouse fibroblasts synthesize prostaglandins in response to bradykinin, thrombin, serum, and the ionophore A23187. These agents activate phospholipases, thereby releasing fatty acids from phospholipids. To examine the phospholipid specificity of the phospholipases activated by bradykinin, thrombin, serum, and A23187, cells were labeled with [14C]arachidonic acid and stimulated with these agents in the presence of delipidated bovine serum albumin. Phospholipid classes were resolved by two-dimensional chromatography on silica gel-coated paper. Only phosphatidylinositol and phosphatidylcholine lost radioactivity upon stimulation. To characterize the fatty acid specificity of the phospholipases, cells were incubated with 14C-labeled stearic, oleic, linoleic, eicosatrienoic, or arachidonic acid and then exposed to the stimuli. Bradykinin, thrombin, and serum caused specific release of radioactivity into the medium only from cells labeled with arachidonic acid or eicosatrienoic acid, whereas A23187 caused release from cells labeled with any one of the five fatty acids. We conclude that bradykinin, thrombin, and serum activate phospholipases that specifically hydrolyze arachidonyl and eicosatrienoyl phosphatidylinositol and phosphatidylcholine, whereas A23187 is less specific activator of phospholipases.
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001095:Arachidonic Acids / Q000378:metabolism; D001769:Blood; D001920:Bradykinin / Q000494:pharmacology*; D000001:Calcimycin / Q000494:pharmacology*; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D003470:Culture Media; D004789:Enzyme Activation; D005347:Fibroblasts; D007700:Kinetics; D008748:Methylcholanthrene / Q000494:pharmacology*; D051379:Mice; D010740:Phospholipases / Q000378:metabolism*; D010743:Phospholipids / Q000096:biosynthesis; D013917:Thrombin / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D001095:Arachidonic Acids; D003470:Culture Media; D010743:Phospholipids; D000001:Calcimycin; D008748:Methylcholanthrene; D010740:Phospholipases; D013917:Thrombin; D001920:Bradykinin