酵母系统表达TPO与E.coli表达TPO N端结构域生物半衰期比较  

The Comparison between Biological Half life of TPO Mature Peptide Expressed in Piochia pastoris and TPO N terminal Domain Expressed in E.coli

作　　者：刘丽[1,2] 田生礼[1,2] 王颖[1,2] 芦兴武[1,2] 孟岩 谢宝树[1,2] 刘立忠[1,2] 冷爱军 

机构地区：[1]空军总医院空军临床分子生物学研究中心 [2]东北师范大学细胞与遗传研究所

出　　处：《中国生物化学与分子生物学报》1998年第6期784-789,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金;全军"九五"医药卫生科研基金

摘　　要：为了探讨血小板生成素（ＴＰＯ）糖基化与其生物半衰期的关系，首先纯化了巴氏毕赤酵母系统分泌表达的人ＴＰＯ成熟肽和大肠杆菌表达的人ＴＰＯＮ端结构域蛋白．纯化产物经Ｗｅｓｔｅｒｎｂｌｏｔ鉴定和活性分析后，两种蛋白以相同浓度或相同活性单位给Ｗｉｓｔｅｒ大鼠尾静脉注射，利用酶联免疫分析（ＥＬＩＳＡ）和ＴＦ１细胞测定不同时间点实验动物血浆中ＴＰＯ浓度或生物学活性，求出两种ＴＰＯ的生物半衰期（ｔ１／２）值．结果显示，以相同浓度和相同活性单位给药，ＴＰＯ成熟肽的ｔ１／２均比其Ｎ端结构域的长，前者分别是后者的１．６５倍和１．２７倍．In order to investigate the relation of thrombopoietin (TPO) glycosylation and biological half life,human thrombopoietin mature peptide expressed by Piochia pastoris and TPO N terminal domain protein expressed by E.coli were purified.After Western blot analysis and activity assay,two kinds of proteins were injected into Wister rats with 50 μg/rat or 5×10 5 U/rat by tail vein injection respectively.TPO level in plasma and biological activity were detected at different time points by means of ELISA and TF1 cell line proliferation.Biological half life ( t 1/2 ) of two kinds of TPO were calculated.The results indicated that the half body clearance of TPO mature peptide was longer than that of TPO N terminal domain protein with the same activity unit and same concentration.The former were 1 65 times and 1 27 times as long as the latter respectively.

关 键 词：血小板生成素 糖基化 生物半衰期 N端结构 

分 类 号：Q513.401[生物学—生物化学] Q592