作者: B K Lam , W F Owen , K F Austen , R J Soberman
DOI: 10.1016/S0021-9258(18)51570-2
关键词: Biosynthesis 、 Extracellular 、 Glutathione 、 Leukotriene C4 、 Incubation 、 Biochemistry 、 Leukotriene A4 、 Intracellular 、 Chromatography 、 Chemistry 、 Dinitrophenyl
摘要: We have examined the requirements for export of leukotriene C4 (LTC4) from cultured human eosinophils. To define saturability and kinetics LTC4 export, eosinophils were interacted with A4 (LTA4) at 37 degrees C, methanolic extracts cell-associated extracellular compartments then analyzed content by reverse phase high performance liquid chromatography on-line monitoring absorbance 280 nm. When LTA4 was added concentrations 0 to 100 microM 10 min amount released extracellularly became constant an concentration 7.5 or greater even though intracellular continued increase. incubated 50 0-60 C held remainder 60-min interval, 54.2 77.3% (n = 3), respectively, total after 15 30 incubation C. Eosinophils 1 h synthesized 290 pmol 3) which approximately half-maximal, all retained intracellularly. utilized time temperature dependence preload both C5 (LTC5) sequentially supplying them specific substrates. With increasing LTC5, there dose-dependent inhibition subsequent release sum glutathionyl leukotrienes remaining constant. In addition, only minimal competition occurred when cells preloaded conjugate 1-chloro-2,4-dinitrobenzene reduced glutathione, S-(dinitrophenyl)glutathione. The criteria saturability, LTC5 release, establish as a novel biochemical step distinct uptake conjugation glutathione synthase form LTC4.