title: Enzymatic formation of prostaglandin D2 by rat basophilic leukemia cells and normal rat mast cells.
abstract: It has been shown that the major cyclooxygenase product in rat basophilic leukemia (RBL-1) cells and in normal rat mast cells is prostaglandin D2. In RBL-1 cells, prostaglandin D2 is isomerase activity was found in the 150 000 X g microsomal pellet as well as the supernatant fraction. Incubation of RBL-1 microsomes with arachidonic acid without cofactors yielded 17.5 +/- 2% prostaglandin E2 and 9.1 +/- 1.4% prostaglandin D2. The cyclooxygenase activity was enhanced (25%) by epinephrine and the addition of reduced glutathione led to a marked increase in prostaglandin D2 synthesis (3-fold). Incubations with arachidonic acid, glutathione and epinephrine gave the maximum conversion to prostaglandin D2, yielding 7 +/- 0.4% prostaglandin E2 and 35.6 +/- 3.5% prostaglandin D2. Incubations with [14C]prostaglandin H2 to bypass cyclooxygenase confirmed the presence and glutathione dependence of the prostaglandin D2 isomerase in the microsomal fraction and also revealed the presence of the same enzyme in the 150 000 X g supernant. In contrast to RBL-1 cells, incubations of microsomes and supernatant from normal rat mast cells with [14C]-arachidonic acid and [14C]prostaglandin H2 localized the prostaglandin D2 isomerase activity in the soluble fraction. Similar to the enzyme in the RBL-1 cells, the mast cell enzyme was glutathione dependent.
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000494:pharmacology; D001491:Basophils / Q000378:metabolism*; D002478:Cells, Cultured; D004837:Epinephrine / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology; D066298:In Vitro Techniques; D019746:Intramolecular Oxidoreductases; D007535:Isomerases / Q000378:metabolism; D007942:Leukemia, Experimental / Q000378:metabolism*; D054834:Lipocalins; D008297:Male; D008407:Mast Cells / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism; D011453:Prostaglandins / Q000096:biosynthesis*; D011457:Prostaglandins D / Q000096:biosynthesis*; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D001095:Arachidonic Acids; D054834:Lipocalins; D011453:Prostaglandins; D011457:Prostaglandins D; D007535:Isomerases; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase; D005978:Glutathione; D004837:Epinephrine