分子克隆和人类55kd瘤亡因子受体的表达
H Loetscher1, Y C Pan, H W Lahm
1Central Research Units, F. Hoffmann-LaRoche LTD, Basel, Switzerland.
Cell
|April 20, 1990
概括
研究人员净化了55kd瘤亡因子 (TNF) 受体,并确定了它的氨基酸序列. 这项工作确定了TNF受体.
科学领域:
- 分子生物学分子生物学
- 免疫学 免疫学 免疫学
- 细胞生物学 细胞生物学
背景情况:
- 瘤亡因子 (TNF) 的两个不同的受体,55 kd和75 kd,在各种细胞上以低水平表达.
- 了解TNF受体的结构和功能对于理解炎症和免疫反应至关重要.
研究的目的:
- 为了净化和表征55kd的TNF受体.
- 确定TNF受体功能的分子基础,并确定潜在的治疗点.
主要方法:
- 从HL60细胞中净化55kd的TNF受体.
- 部分氨基酸序列的确定.
- 反转录酶-聚合酶连锁反应 (RT-PCR) 使用退化的寡核酸原始剂来放大cDNA片段.
- 使用放大片段作为探针对人类胎盘cDNA库的查.
- 克隆cDNA在COS-1细胞和baculovirus系统中的表达,以评估TNF结合性质.
主要成果:
- 确定了55kd TNF受体的部分氨基酸序列.
- 一个66bpcDNA片段被放大并用于识别人类胎盘cDNA库中的重叠克隆.
- 全长的cDNA预测了一个455氨基酸TNF受体蛋白,具有不同的功能域 (领导,细胞外,膜外,细胞内).
- 表达的TNF受体赋予了与原生受体相当的TNF结合特性.
- 在55kd TNF受体和NGF受体细胞外域之间观察到显著的序列同质性.
结论:
- 该研究成功净化和表征了55kd TNF受体,阐明了其分子结构.
- 这些发现为了解TNF受体信号通路及其在疾病中的作用提供了基础.
- 与NGF受体的同质性表明潜在的共享信号机制或进化关系.
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Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
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Such genes that act...
Loss of Tumor Suppressor Gene Functions
Tumor suppressor genes are normal genes that can slow down cell division, repair DNA mistakes, or program the cells for apoptosis in case of irreparable damage. Hence, they play an essential role in preventing the proliferation of damaged cells.
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