Ohnishi H, Kosuzume H, Kitamura Y, Yamaguchi K, Nobuhara M, Suzuki Y, Yoshida S, Tomioka H, Kumagai A
Prostaglandins. 1980 Oct;20(4):655-66. doi: 10.1016/0090-6980(80)90106-9.
To elucidate the chemical structure of slow-reacting substance of anaphylaxis from rat (SRS-A rat), SRS-A rat were purified by the method of Orange with modification using DEAE-Sephadex A-25 chromatography. Ultraviolet absorption spectrum of purified SRS-A rat indicated the presence of conjugated triene. Arylsulfatase B degradation products and HCl degradation products were subjected to analysis by a gas chromatography and mass spectrometry and a thin layer chromatography. Products obtained by arylsulfatase b catalysis contained 5,6-dihydroxy-7,9,11,14-eicosatetraenoic acid. HCl degradation products showed the presence of glycine, glutamic acid and cysteic acid. Furthermore, the analysis of anhydrous hydrazine degradation products of SRS-A rat and of HCl hydrolyzed products of dinitrophenylated SRS-A rat revealed the presence of glycine at C-terminal and glutamine acid at N-terminal. The study of the substrate specificity of arylsulfatase B against various materials including SRS-A rat suggested the presence of sulfone in SRS-A rat. The molecular ion peak of SRS-A rat sodium salt was observed at m/e 680 in field desorption mass spectrum of SRS-A rat. On the basis of these data, we identified the structure of SRS-A rat as [gamma]glutamyl-4(5-hydroxy-7,9,11,14-eicosatetraenoic acid-6-yl)-4,4-dioxyocysteinyl] glycine.
为阐明大鼠过敏反应慢反应物质（SRS-A大鼠）的化学结构，采用改良的Orange法，利用DEAE-葡聚糖A-25柱色谱对SRS-A大鼠进行纯化。纯化后的SRS-A大鼠的紫外吸收光谱表明存在共轭三烯。对芳基硫酸酯酶B降解产物和盐酸降解产物进行气相色谱-质谱分析和薄层色谱分析。芳基硫酸酯酶b催化得到的产物含有5,6-二羟基-7,9,11,14-二十碳四烯酸。盐酸降解产物显示存在甘氨酸、谷氨酸和半胱氨酸。此外，对SRS-A大鼠的无水肼降解产物和二硝基苯基化SRS-A大鼠的盐酸水解产物的分析表明，C末端存在甘氨酸，N末端存在谷氨酸。对芳基硫酸酯酶B针对包括SRS-A大鼠在内的各种物质的底物特异性研究表明SRS-A大鼠中存在砜。在SRS-A大鼠的场解吸质谱中，观察到SRS-A大鼠钠盐的分子离子峰为m/e 680。基于这些数据，我们将SRS-A大鼠的结构鉴定为[γ]谷氨酰-4(5-羟基-7,9,11,14-二十碳四烯酸-6-基)-4,4-二氧代半胱氨酰]甘氨酸。
