Human tumor necrosis factor-alpha receptor. Purification by immunoaffinity chromatography and initial characterization.
Human tumor necrosis factor-alpha receptor. Purification by immunoaffinity chromatography and initial characterization.
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人肿瘤坏死因子-α受体。
DOI：
10.1016/s0021-9258(18)37395-2
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
B B Aggarwal
中科院分区：
文献类型：
--
作者：
G B Stauber;R. Aiyer;B B Aggarwal
从人组织细胞淋巴瘤细胞系U937的亚克隆中分离出人肿瘤坏死因子-α（TNF-α）的受体。这些细胞表现出单一类别的高亲和力受体（Kd = 0.51 +/- 0.25 nM），平均密度为55，000 +/-5，000个结合位点/细胞。在用去污剂溶解后，受体保留其结合游离TNF-α的能力，但不能结合固定在各种固体支持物上的TNF-α。对于受体纯化，通过双功能交联试剂乙二醇双（琥珀酰亚胺基琥珀酸酯）或3，3-二硫代双（磺基琥珀酰亚胺基丙酸酯）将125 I-TNF-α共价连接至完整细胞上的受体。然后用非离子去污剂Triton X-100溶解细胞，离心澄清的上清液通过由TNF-α抗血清制备的IgG-琼脂糖柱。通过制备型十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进一步纯化来自抗体柱的富含受体的级分。这两个步骤一起提供了TNF-α受体的约165，000倍纯化。当通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测时，通过该方法获得的TNF-α受体-配体复合物的亚基分子量为100，000 +/-5，000，但在凝胶过滤时，复合物迁移的表观分子量为480，000 +/-32，000。然而，当在不存在配体的情况下进行凝胶过滤时，受体显示出65，000 +/-32，000的分子量。的受体的其他特性进行了讨论。
The receptor for human tumor necrosis factor-alpha (TNF-alpha) was isolated from a subclone of the human histiocytic lymphoma cell line U937. These cells exhibit a single class of high affinity receptors (Kd = 0.51 +/- 0.25 nM) with an average density of 55,000 +/- 5,000 binding sites/cell. After solubilization with detergent, the receptor retained its ability to bind free TNF-alpha but failed to bind to TNF-alpha immobilized on various solid supports. For receptor purification, 125I-TNF-alpha was covalently attached to the receptor on intact cells by the bifunctional cross-linking reagents ethylene glycolbis(succinimidylsuccinate) or 3,3-dithiobis(sulfosuccinimidylpropionate). The cells were then solubilized with the nonionic detergent Triton X-100, and the supernatants, clarified by centrifugation, were passed over an IgG-Sepharose column prepared from TNF-alpha antiserum. The receptor-rich fraction from the antibody column was further purified by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These two steps together provided approximately 165,000-fold purification of the TNF-alpha receptor. The TNF-alpha receptor-ligand complex obtained by this method had a subunit molecular weight of 100,000 +/- 5,000 when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis but on gel filtration the complex migrated with an apparent molecular weight of 480,000 +/- 32,000. However, the receptor showed a molecular weight of 65,000 +/- 32,000 when gel filtration was performed in the absence of ligand. Additional characteristics of the receptor are discussed.
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影响因子：
3.9
作者：
Pal, Nabanita;Kim, Taeyeon;Park, Jae-Seo;Cho, Eun-Bum
通讯作者：
Cho, Eun-Bum
对生物合成标记的 Leydig 肿瘤细胞中的促黄体激素/绒毛膜促性腺激素受体进行免疫沉淀。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kim,IC;Ascoli,M;Segaloff,DL
通讯作者：
Segaloff,DL
人和鼠肿瘤坏死因子的物种特异性。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Smith,RA;Kirstein,M;Fiers,W;Baglioni,C
通讯作者：
Baglioni,C
DOI：
10.1073/pnas.82.24.8667
发表时间：
1985-12-01
影响因子：
11.1
作者：
GAMBLE, JR;HARLAN, JM;VADAS, MA
通讯作者：
VADAS, MA
干扰素-γ增强肿瘤坏死因子细胞受体的表达。
DOI：
--
发表时间：
1986
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Tsujimoto,M;Yip,YK;Vilcek,J
通讯作者：
Vilcek,J