大鼠尿激肽释放酶的纯化及化学研究。

Purification and chemical studies on rat urinary kallikrein.

作者信息

Porcelli G, Marini-Bettòlo G B, Croxatto H R, Di Iorio M

出版信息

Ital J Biochem. 1975 May-Aug;24(3-4):175-87.

Abstract

A highly purified kallikrein was obtained from rat urine by chromatography on DE-32 cellulose, affinity chromatography on Bio-gel P-200-Aprotinin and gel filtration over Sephadex G-100 coarse and superfine. A molecular weight of 32,000 by sodium dodecyl sulfate polyacrylamide disc gel electrophoresis was estimated. The aminoacid composition and the esterase activity of the purified material were determined. Biological characterization of the purified kallikrein was tested by liberation of a kinin from rat plasma kininogen, by direct action on the isolated rat uterus and by the lowering of rat arterial pressure after intravenous injection of the enzyme. The preparation of insoluble derivative of Aprotinin is described herein. The polymer used as insoluble support (Bio-gel P-200) was before changed to its corresponding azide, which reacts with Aprotinin; the product maintained the binding property of the Aprotinin with urinary kallikrein.

摘要

通过在DE - 32纤维素上进行色谱分离、在Bio - gel P - 200 -抑肽酶上进行亲和色谱分离以及在Sephadex G - 100粗颗粒和超细颗粒上进行凝胶过滤，从大鼠尿液中获得了高度纯化的激肽释放酶。通过十二烷基硫酸钠聚丙烯酰胺圆盘凝胶电泳估计其分子量为32,000。测定了纯化物质的氨基酸组成和酯酶活性。通过从大鼠血浆激肽原中释放激肽、对离体大鼠子宫的直接作用以及静脉注射该酶后降低大鼠动脉血压，对纯化的激肽释放酶进行了生物学特性测试。本文描述了抑肽酶不溶性衍生物的制备。用作不溶性支持物（Bio - gel P - 200）的聚合物预先转化为其相应的叠氮化物，该叠氮化物与抑肽酶反应；产物保留了抑肽酶与尿激肽释放酶的结合特性。