尿激酶短肽抑制剂的初步研究

Selection of peptidic inhibitors of urokinase from a phage peptide library

以尿激酶为目标蛋白,在噬菌体表面展示六肽库中对尿激酶的短肽类抑制剂进行了三轮特异性筛选.提高噬菌体与尿激酶的比例及缩短作用时间从而提高筛选压力后,与尿激酶亲和结合的噬菌体得到富集.通过对第三轮筛选到的重组噬菌体的DNA序列分析,获得一组相对保守的肽序列.相应的合成短肽NEPKAN和VSPKVL对尿激酶的抑制常数分别为32.5μmol/L和88.6μmol/L.

The urokinase peptide inhibitors were screened in a phage display 6-mer peptide library by three rounds of selection basically according to the method described by Smith. To improve the screening effectiveness and specificity, we designed a novel screening method in order to remove most of urokinase substrates from the peptide library. The potential inhibitors of urokinase were riched. After sequencing the clones, several related peptides were determined. Two 6-mer peptides were synthesized and their inhibitory constants to urokinase were determined to be (88.6 μmol/L) and (32.5 μmol/L,) respectively.