摘要
以肾衰病人尿为材料,通过碱变性、酸变性和热变性除去部分杂质和尿蛋白。然后用羧甲基-木瓜蛋白酶-琼脂糖4B亲和层析,最后用多缓冲剂聚焦层析,把人的巯基蛋白酶抑制肽C(human cysteine proteinase inhibitor C,简称 hCPI_C)和其他的hCPI_C分开,制备得高纯度、高活性的hCPI_C。在SDS-PAGE中为均一条带,每mg蛋白可抑制50单位木瓜蛋白酶活性的50-63％。
Human cysteine proteinase inhibitor C has been purified by precipitating part of urine protein with acid, base and heat denaturation followed by, Cm-papain-sepharose 4B affinity chromatography, and finally using polybuffer exchanger 94 chromatofocusing to separate hCPIc from other hCPI.The preparation is high in purity with high inhibitory activity against papain.The hCPIc prepared exhibits on ly one band of protein on SDS-PAGE 1mg of this protein preparation is able to inhibits 50-60% of the proteinase activity in 50 BAEE units of papain.
基金
国家自然科学基金
关键词
聚焦层析
巯基蛋白酶
抑制肽C
Human Cysteine Proteinase Inhibitor C
Chromatofocusing
Affinity Chro-matography
Cathepsin
