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Fusion expression and purification of discoidin domain receptor 2 extracellular domain in Pichia pastoris and its functional analysis
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 14
- DOI:
- 10.3321/j.issn:1000-2790.2007.14.001
- Key Word:
- 盘状结构域受体2;胞外区;基因表达;阻断剂
Abstract： 目的:盘状结构域受体2(discoidin domain receptor2,DDR2)是一种与肿瘤细胞转移相关的蛋白酪氨酸激酶,研究DDR2在类风湿性关节炎(rheumatoid arthritis,RA)软骨破坏和肿瘤转移中的作用.方法:在毕赤酵母中表达DDR2胞外区片段(不含信号肽和跨膜区,命名为DR),并将所表达的蛋白进行纯化和功能鉴定,以备将来用作DDR2的特异性阻断剂.结果:获得了两株较高表达His-DR融合蛋白的毕赤酵母克隆;其表达的蛋白质中可溶性部分约占全部融合蛋白的18%;经Ni-NTA(nitric-tri-acetic acid)柱纯化后,获得了纯度约90%以上的可溶性His-DR融合蛋白;竞争结合抑制实验显示,His-DR具有阻断和细胞表面天然DDR2受体结合的功能.结论:所表达的融合蛋白His-DR具有抑制天然DDR2功能的作用.
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