Leukotriene C4 and D4 formation by particulate enzymes.
The homogenate of rat basophilic leukemia cells, when incubated with arachidonic acid, glutathione, and calcium, formed 3 isomers of 5,12-dihydroxyeicosatetraenoic acid and 2 isomers of 5,6-dihydroxyeicosatetraenoic acid, as well as leukotriene (LT) C4 and D4. The products were identified by high pressure liquid chromatography, ultraviolet spectral analysis, co-migration with standards, bioassay, and gas chromatography-mass spectrometry. The enzymes responsible for the formation of LTC4 and LTD4 from LTA4 were found in the 10,000 x g pellet and, therefore, appear to be particulate. The possibility that these enzymes are bound to the cell membrane suggest that the formation of these leukotrienes might be important in the basophil and mast cells release reaction.