大鼠胃激肽释放酶：其纯化及性质

Rat stomach kallikrein: its purification and properties.

作者信息

Uchida K, Yokoshima A, Niinobe M, Kato H, Fujii S

出版信息

Adv Exp Med Biol. 1979;120A:291-303. doi: 10.1007/978-1-4757-0926-1_28.

Abstract

From rat stomach, kallikrein was purified by chromatographies on columns of p-aminobenzamidine-Sepharose, DEAE-Sephadex A-50 and Sephadex G-150 and by isoelectric focusing, measuring its activities to hydrolyse prolylphenylalanyl-arginine-4-methyl-coumarine amide (Pro-Phe-Arg-MCA) and to release kinin from rat heated-plasma. The purified stomach kallikrein showed a single band on Disc electrophoresis at pH 7.0. The molecular weight of the kallikrein was calculated to be 29,000 by gel-filtration on a column of Sephadex G-50. The kallikrein was stable between pH 6 and 11 and hydrolysed Pro-Phe-Arg-MCA optimally at pH 11.0. The Pro-Phe-Arg-MCA hydrolysing activity of rat stomach kallikrein was inhibited by DFP and Trasylol, but not by trypsin inhibitors from soyabean, limabean and ovomucoid. These properties of rat stomach kallikrein was clearly distinguishable from those of partially purified rat plasma kallikrein, but similar properties to other glandular kallikreins from other species. From these results, it was concluded that kallikrein is present in rat stomach, which can be classified into glandular kallikrein.

摘要

从大鼠胃中提取激肽释放酶，通过对氨基苯甲脒-琼脂糖柱、二乙氨基乙基-葡聚糖A-50柱和葡聚糖G-150柱进行色谱分离以及等电聚焦法进行纯化，并测定其水解脯氨酰苯丙氨酰精氨酸-4-甲基香豆素酰胺（Pro-Phe-Arg-MCA）的活性以及从大鼠加热血浆中释放激肽的活性。纯化后的胃激肽释放酶在pH 7.0的圆盘电泳中呈现单一谱带。通过在葡聚糖G-50柱上进行凝胶过滤，计算出激肽释放酶的分子量为29,000。该激肽释放酶在pH 6至11之间稳定，在pH 11.0时对Pro-Phe-Arg-MCA的水解活性最佳。大鼠胃激肽释放酶的Pro-Phe-Arg-MCA水解活性受到二异丙基氟磷酸酯（DFP）和抑肽酶的抑制，但不受来自大豆、利马豆和卵类粘蛋白的胰蛋白酶抑制剂的抑制。大鼠胃激肽释放酶的这些特性与部分纯化的大鼠血浆激肽释放酶的特性明显不同，但与其他物种的其他腺体激肽释放酶的特性相似。从这些结果可以得出结论，激肽释放酶存在于大鼠胃中，可归类为腺体激肽释放酶。