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天花粉蛋白的定点聚乙二醇修饰
引用本文:何贤辉,聂慧玲,邵鹏柱,柯一保,谭兆祥.天花粉蛋白的定点聚乙二醇修饰[J].中国生物化学与分子生物学报,1999,15(6):924-927.
作者姓名:何贤辉  聂慧玲  邵鹏柱  柯一保  谭兆祥
作者单位:中国科学院上海细胞生物学研究所!上海2000311(何贤辉,柯一保),中国科学院上海细胞生物学研究所!上海2000311(聂慧玲),香港中文大学生物化学系!香港(邵鹏柱),香港中文大学生理学系!香港(谭兆祥)
摘    要: 用一种定点修饰天花粉蛋白(trichosanthin,TCS)的方法,将聚乙二醇(PEG)偶联到预先选定的位点.利用nTCS无半胱氨酸(Cys)残基这一特点,通过定点突变将一个Cys残基引入TCS以取代第7位的丝氨酸(Ser)残基.然后,与巯基反应的PEG-m aleim ide 即可偶联到新引入的Cys 残基上.经纯化得到均一的PEG-TCS复合物,在SDS-PAGE上显示一条区带,表观分子量为38 kD.复合物的体外致核糖体失活活性降低了6倍,但其体内引产活性与nTCS相同.定点PEG修饰方法为改造TCS提供了新途径.

关 键 词:天花粉蛋白  定点突变  聚乙二醇修饰  致核糖体失活活性
收稿时间:1999-12-20

Site-Directed Polyethylene Glycol Modification of Trichosanthin
HE Xianhui,NIE Huiling,SHAW Pangchui ,KE Yibao,TAM Siucheung.Site-Directed Polyethylene Glycol Modification of Trichosanthin[J].Chinese Journal of Biochemistry and Molecular Biology,1999,15(6):924-927.
Authors:HE Xianhui  NIE Huiling  SHAW Pangchui  KE Yibao  TAM Siucheung
Institution:(Shanghai Institute of Cell Biology,Academia Sinica,Shanghai 200031;Departments of 1) Biochemistry and 2) Physiology, The Chinese University of Hong Kong,
Abstract:The site directed polyethylene glycol(PEG)modification(PEGylation)had been establishied to couple PEG to a pre selected site on trichosanthin(TCS)which in its natural form had no Cys residue.By site directed mutagenesis,a Cys residue can be introduced to replace the 7th residue Ser and PEG was then coupled to this newly created Cys by PEG maleimide.The PEG TCS conjugate was purified to homogeneity and showed a single band on SDS PAGE with an apparent molecular weight of 38 kD.Its ribosome inactivating activity in vitro was reduced by 6 fold,whereas the abortifacient activity in vivo was comparable to nTCS.The site directed PEGylation has provided a new approach for reconstructing TCS.
Keywords:Trichosanthin  Site  directed mutagenesis  PEGylation  Ribosome-inactivating activity
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