分子克隆和表达一个受体的人类瘤亡因子
T J Schall1, M Lewis, K J Koller
1Department of Molecular Biology, Genentech, Inc., South San Francisco, California 94080.
Cell
|April 20, 1990
概括
研究人员在癌症患者血清中发现了一种可溶性瘤亡因子 (TNF) 结合蛋白. 这种蛋白质很可能来自TNF受体 (TNF-R),TNF信号传递至关重要的细胞表面分子.
科学领域:
- 免疫学 免疫学 免疫学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 瘤亡因子 (TNF) 是一种关键的细胞因子,与炎症和免疫有关.
- 在癌症患者血清中检测到可溶性TNF结合蛋白,但其来源尚不清楚.
研究的目的:
- 从血清中净化和表征一种人类TNF结合蛋白.
- 确定编码这种结合蛋白的基因并阐明其与TNF受体的关系.
主要方法:
- 蛋白质净化和部分测序.
- 寡核酸导向的突变发生和cDNA克隆.
- 哺乳动物细胞转染和受体结合试验.
- 北方斑点分析.
主要成果:
- 从癌症患者的血清中净化了一种同质的TNF结合蛋白.
- 分离了编码TNF受体 (TNF-R) 的cDNA克隆.
- TNF-R是一种415氨基酸的跨膜蛋白,与NGF受体具有同质性.
- 转染细胞表达TNF-R,特异结合TNF-α,这种结合被未标记的TNF-α和TNF-β抑制.
- 北方分析显示,在各种细胞类型中,单一的TNF-RmRNA物种存在.
结论:
- 在人血清中发现的可溶性TNF结合蛋白可能是TNF受体的蛋白质分解释放的片段.
- 这一发现为癌症患者中可溶性TNF结合蛋白的存在提供了分子基础,并对TNF介导信号传递有影响.
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