Warwas M, Gburek J, Osada J, Gołab K
Department of Pharmaceutical Biochemistry, Medical Academy, Wrocław, Poland.
Acta Biochim Pol. 1995;42(3):351-6.
It is the second peptidase inhibitor, after ovostatin, which showing the same antipapain activity in egg white in different avian species implies differences in amino-acid sequences. Cystatin from duck egg white was purified by carboxymethylpapain affinity chromatography and size-exclusion HPLC. The purified inhibitor which showed partial identity in the immunodiffusion test with chicken egg white cystatin, had an apparent molecular mass of 9.3 kDa as determined by SDS/PAGE. IEF analysis revealed five molecular forms of pI in the range 7.8-8.4. The obtained cystatin was neither glycosylated nor phosphorylated as it is in the case of chicken cystatin. The determined Ki (0.005 +/- 0.001 nM) was similar to that reported for human and chicken cystatin C.
它是继抑卵素之后的第二种肽酶抑制剂,在不同鸟类物种的蛋清中表现出相同的抗木瓜蛋白酶活性,这意味着氨基酸序列存在差异。通过羧甲基木瓜蛋白酶亲和色谱和尺寸排阻高效液相色谱法对鸭蛋清中的半胱氨酸蛋白酶抑制剂进行了纯化。纯化后的抑制剂在免疫扩散试验中与鸡蛋清半胱氨酸蛋白酶抑制剂有部分一致性,通过SDS/PAGE测定其表观分子量为9.3 kDa。IEF分析揭示了五种等电点在7.8 - 8.4范围内的分子形式。所获得的半胱氨酸蛋白酶抑制剂既没有糖基化也没有磷酸化,这与鸡半胱氨酸蛋白酶抑制剂的情况相同。测定的Ki(0.005±0.001 nM)与报道的人和鸡半胱氨酸蛋白酶抑制剂C的Ki相似。