Enzymatic study to characterize the slow reacting substance of anaphylaxis (SRS-A) and leukotrienes.
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概要
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Slow reacting substance of anaphylaxis (SRS-A) has been shown to be one of the major mediators in hypersensitive reactions and to be composed of leukotriene (LT) C<SUB>4</SUB>, LTD<SUB>4</SUB> and LTE<SUB>4</SUB>. In the present study, we examined the properties of SRS-A released from sensitized guinea pig lungs by antigen and SRS released from rat peritoneal exudate cells and from human leucocytes by ionophore A23187 (0.5 and 0.2 μg/ml, respectively). By the incubation with SRS-A, SRS and LTs with arylsulfatase (type V) in pH 5.7 buffered solution at 37°C for 30 min, SRS-A and LTD<SUB>4</SUB> were greatly inactivated and rat SRS was slightly inactivated, but human SRS and LTC<SUB>4</SUB> were not inactivated at all. The same results were obtained when aminopeptidase was used in place of arylsulfatase. Moreover, when SRS-A, LTC<SUB>4</SUB> and LTD<SUB>4</SUB> were incubated with 0.02 mg/ml of γ-glutamyltranspeptidase (γ-GTP) pH 8.0 buffered solution at 37°C for 30 min, the activities of SRS-A and LTD<SUB>4</SUB> were slightly decreased, but those of SRS and LTC<SUB>4</SUB> were obviously potentiated. On the other hand, incubation with a large amount of γ-GTP (0.2 mg/ml) a dose at which this enzyme preparation showed clear aminopeptidase activity, SRS-A, SRS, LTC<SUB>4</SUB> and LTD<SUB>4</SUB> were obviously inactivated. In addition, we found a peak of LTD<SUB>4</SUB> in guinea pig SRS-A, that of LTC<SUB>4</SUB> in human SRS, and that of LTC<SUB>4</SUB> in rat SRS on high performance liquid chromatograms. From these results, we demonstrated that guinea pig lung SRS-A is mainly composed of LTD<SUB>4</SUB>, human leukocyte SRS is mainly LTC<SUB>4</SUB>, and rat peritoneal SRS is composed of both LTC<SUB>4</SUB> and LTD<SUB>4</SUB>. The inactivation of LTD<SUB>4</SUB> and SRS-A by arylsulfatase may be due to aminopeptidase contamination in the enzyme preparation.
著者
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Kojima Masami
Central Research Laboratory Kissei Pharmaceutical
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Ujiie Arao
Central Research Laboratories Kissei Pharmaceutical Co. Ltd.
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Tsutsumi Naoyuki
Central Research Laboratories Kissei Pharmaceutical Co. Ltd.
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IKEDA Shigeru
Central Research Laboratories, Kissei Pharmaceutical Cc., Ltd.
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KOMATSU Hidetada
Central Research Laboratories, Kissei Pharmaceutical Cc., Ltd.
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