半夏胰蛋白酶抑制剂的纯化与部分性质研究

Purification and Some Properties of a Trypsin Inhibitor from Rhizoma pinelliae

应用以胰蛋白酶为配体的亲和层析法,从生半夏蛋白粗提液的40%(NH4)2SO4(M/V)沉淀中分离纯化出一种胰蛋白酶抑制剂(Rhizoma pinelliae trypsin inhibitor,RPTI),经SDS-PAGE检测呈现单一条带,分子量为14kD左右,其N-端6个氨基酸残基顺序为DPVVDG。研究表明,其对胰蛋白酶的质量抑制比为1∶4.72左右,对人低分化胃腺癌细胞系(BGC-823)的细胞增殖具有抑制作用,IC50值为121.53μg/mL。

A Rhizoma pinelliae trypsin inhibitor（ RPTI ） was purified from the 40% （ NH4 ） 2 S04 （M/V） sediments of the crude extracted protein of pinelliae by the affinity chromatography of trypsin-Sepharose CL 4B. The purified RPTI showed a single band on SDS-PAGE gel with an estimated molecular weight of 14 kD, and the N-terminal sequence was DPVVDG. The quality inhibiting ratio of RFrI to trypsin was about 1： 4.72. RPTI could markedly inhibit the proliferation of BGC-823 cells in vitro,and the IC5o value was about 121.53μg/mL.