Creasey A A, Yamamoto R, Vitt C R
Proc Natl Acad Sci U S A. 1987 May;84(10):3293-7. doi: 10.1073/pnas.84.10.3293.
We compared the molecular structure of the receptor to human recombinant tumor necrosis factor (HurTNF) on cells of different tissue origin that differ in their response to one of the known activities of TNF. We studied tumor cell lines that respond to the cytotoxic action of TNF and resistant variants that bind TNF, normal cell lines that are stimulated to proliferate by TNF and those that are not affected by TNF, and peripheral blood granulocytes whose activation is also augmented by TNF. Using 125I-labeled HurTNF, we found that it bound mainly to four cellular polypeptides (138, 90, 75, and 54 kDa), three of which were found in every cell type examined and one (138 kDa) that was observed only in a human breast carcinoma cell line (MCF-7) that is highly responsive to the cytotoxic action of TNF. The 138-kDa polypeptide was not found in resistant variants of MCF-7 that bind TNF. In contrast to the other polypeptides, the 138-kDa protein was detected 30 min after incubation at 4 degrees C, as compared to 5 min. Scatchard analysis and cross-linking data suggest a model for the TNF receptor structure whereby the receptor is composed of noncovalently linked membrane-bound polypeptides that bind TNF with high affinity (Kd, 0.05-0.8 X 10(-9) M) with the 138-kDa protein being the least abundant and/or even absent in most cells.
我们比较了不同组织来源细胞上该受体与重组人肿瘤坏死因子（HurTNF）的分子结构，这些细胞对TNF已知活性之一的反应有所不同。我们研究了对TNF细胞毒性作用有反应的肿瘤细胞系以及能结合TNF的抗性变体、受TNF刺激而增殖的正常细胞系和不受TNF影响的正常细胞系，还有激活也会因TNF增强的外周血粒细胞。使用125I标记的HurTNF，我们发现它主要与四种细胞多肽（138、90、75和54 kDa）结合，其中三种在每种检测的细胞类型中都能找到，另一种（138 kDa）仅在对TNF细胞毒性作用高度敏感的人乳腺癌细胞系（MCF-7）中观察到。在能结合TNF的MCF-7抗性变体中未发现138-kDa多肽。与其他多肽不同，138-kDa蛋白在4℃孵育30分钟后被检测到，而其他多肽在5分钟后就能检测到。Scatchard分析和交联数据提示了TNF受体结构的一种模型，即该受体由非共价连接的膜结合多肽组成，这些多肽以高亲和力（Kd，0.05 - 0.8×10(-9) M）结合TNF，其中138-kDa蛋白在大多数细胞中含量最少甚至不存在。
