Mouse peritoneal macrophages release leukotriene C in response to a phagocytic stimulus.
Mouse peritoneal macrophages that had ingested zymosan particles released a polar metabolite of arachidonic acid possessing slow-reacting substance activity in the guinea pig ileum assay. The metabolite was purified by solvent extraction, Sephadex G-25 column chromatography. The purified metabolite absorbed light at 280 nm and contained a free amino group. When macrophages were preincubated overnight with [3H]arachidonic acid, [35H]cysteine, or [14C]glutamic acid, each radiolabel was incorporated into the compound. Direct amino acid analysis revealed glycine, glutamic acid, and cysteine at molar ratios of 0.97:1.00:0.82. The above data were consistent with the structure of leukotriene C, an adduct of arachidonic acid and glutaathione. Quantification of the leukotriene C based on incorporation of [3H]arachidonic acid or amino acid analysis indicated that 6 X 10(7) macrophages (3.6 mg of cell protein) released 7.5 nmol after a maximal phagocytic stimulus. The purified leukotriene C had a slow reacting substance activity of 11,500 units/nmol (1 unit has the activity of 5 ng of histamine in a guinea pig ileum contraction assay).
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001202:Ascitic Fluid / Q000166:cytology; D005260:Female; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D010587:Phagocytosis; D013189:SRS-A / Q000302:isolation & purification / Q000378:metabolism*
D000596:Amino Acids; D013189:SRS-A