Pace-Asciak C R, Klein J, Lombard S, Torchia J, Rokach J
Biochim Biophys Acta. 1985 Aug 22;836(1):153-6. doi: 10.1016/0005-2760(85)90231-0.
[3H]Leukotriene A4 was incubated with various subcellular fractions of rat liver homogenates. After solvent extraction and purification on C18 Sep-Pak cartridges, tritiated products migrating on reversed-phase HPLC with authentic unlabelled leukotriene C4, D4 and B4 were observed. The identity of leukotriene C4 was confirmed through enzymatic conversion into D4 by gamma-glutamyl transpeptidase as well as by bioassay on the rat stomach fundus after HPLC purification. The contractile response to the extracted material was blocked by the SRS antagonist, FPL 55712. Leukotriene B4 synthesis was located in the 100 000 X g supernatant, while C4 synthesis was present in the corresponding pellet. Leukotriene C4 formation was enhanced when reduced glutathione was supplemented in the incubation medium. These results demonstrate the presence in rat liver of various enzymatic steps in leukotriene A4 catabolism.
将[3H]白三烯A4与大鼠肝脏匀浆的各种亚细胞组分一起孵育。经溶剂萃取并在C18 Sep-Pak柱上纯化后，观察到氚化产物在反相高效液相色谱上与未标记的白三烯C4、D4和B4一同迁移。白三烯C4的身份通过γ-谷氨酰转肽酶将其酶促转化为D4以及在高效液相色谱纯化后对大鼠胃底进行生物测定得以确认。对提取物的收缩反应被SRS拮抗剂FPL 55712阻断。白三烯B4的合成位于100000×g上清液中，而C4的合成则存在于相应的沉淀中。当在孵育培养基中补充还原型谷胱甘肽时，白三烯C4的形成会增强。这些结果证明大鼠肝脏中存在白三烯A4分解代谢的各种酶促步骤。
