Characterization of the two major species of slow reacting substance from rat basophilic leukemia cells as glutathionyl thioethers of eicosatetraenoic acids oxygenated at the 5 position. Evidence that peroxy groups are present and important for spasmogenic activity.
The most prominent slow reacting substance from rat basophilic leukemia cells (type I) was characterized by radiochemical, chemical and physical methods and shown to contain a C20 unsaturated fatty acid oxygenated at the 5 position and a sulfur containing side chain in thioether linkage at the 6 position. Its spasmogenic action on guinea pig ileal muscle was largely inactivated under reducing conditions which suggested that a peroxy group was present and important for contractile activity. This was supported by ferrous thiocyanate analysis. The peroxy group is almost certainly at the 5 position, probably in the form of a peroxy ester or hydroperoxide. Based on amino acid hydrolysis (0.85 moles of glycine and 0.30 moles of glutamic acid per mole SRS), the sulfur containing side chain is apparently a mixture of glutathione and cysteinyl-glycine, but by chromatography the side chain is predominantly glutathione and the low yield of glutamic acid may be due to complexing of its alpha COOH group in a peroxy ester linkage. The fatty acid moiety has 3 conjugated double bonds, probably at the 7,8, 9,10 and 11,12 positions. Type II SRS, the second major species, differs in that the sulfur containing side chain is linked at the 12 or 13 position and is almost certainly glutathione and in the failure of alkaline borohydride to produce inactivation. These observations strongly implicate the lipoxygenase pathway in slow reacting substance biosynthesis.
D015117:5,8,11,14-Eicosatetraynoic Acid / Q000031:analogs & derivatives / Q000032:analysis*; D000818:Animals; D012898:Autacoids / Q000032:analysis / Q000097:blood / Q000494:pharmacology*; D001491:Basophils / Q000032:analysis*; D055598:Chemical Phenomena; D002621:Chemistry; D005231:Fatty Acids, Unsaturated / Q000032:analysis*; D006168:Guinea Pigs; D007942:Leukemia, Experimental / Q000097:blood*; D009119:Muscle Contraction / Q000187:drug effects; D010545:Peroxides / Q000032:analysis; D051381:Rats; D013455:Sulfur / Q000032:analysis
D012898:Autacoids; D005231:Fatty Acids, Unsaturated; D010545:Peroxides; D015117:5,8,11,14-Eicosatetraynoic Acid; D013455:Sulfur