Structure of the murine erythropoietin receptor complex. Characterization of the erythropoietin cross-linked proteins.
Structure of the murine erythropoietin receptor complex. Characterization of the erythropoietin cross-linked proteins.
复制标题
鼠促红细胞生成素受体复合物的结构。
DOI：
10.1016/s0021-9258(18)54507-5
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Sylvie Gisselbrecht
中科院分区：
文献类型：
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作者：
P. Mayeux;Catherine Lacombe;Nicole Casadevall;S. Chrétien;I. Dusanter;Sylvie Gisselbrecht
使用针对克隆的促红细胞生成素受体链的细胞内部分的抗体研究了鼠促红细胞生成素受体的结构。这些抗体从Triton X-100溶解的细胞中沉淀出促红细胞生成素受体复合物。当复合物交联的二琥珀酰亚胺辛二酸酯，85-和100-kDa的促红细胞生成素交联的蛋白质先前描述的免疫沉淀。然而，当复合物在免疫沉淀之前变性和还原时，这些蛋白质没有沉淀。使用1-乙基3-（3-二甲基氨基丙基）碳二亚胺，我们观察到红细胞生成素交联的蛋白质的66 kDa的除了100-和85-kDa的蛋白质。只有66-kDa的促红细胞生成素交联蛋白的免疫沉淀后，变性和还原的复合物的抗受体抗体。因此，我们的研究结果表明，85-和100-kDa的蛋白质先前证明的交联与受体的克隆链，形成一个多聚体复合物，但这些蛋白质似乎免疫无关的克隆链。我们观察到，减少能够交联氨基的分子的长度会降低与100-kDa蛋白质的交联效率，并且只有85-kDa蛋白质使用1，5-二氟-2，4-二硝基苯与促红细胞生成素交联。这些结果表明，85-和100-kDa的蛋白质结合的红细胞生成素分子的受体相对略有不同的位置。
The structure of the murine erythropoietin receptor was studied using antibodies against the intracellular part of the cloned erythropoietin receptor chain. These antibodies precipitated erythropoietin-receptor complexes from Triton X-100-solubilized cells. When the complexes were cross-linked by disuccinimidyl suberate, the 85- and 100-kDa erythropoietin-cross-linked proteins previously described were immunoprecipitated. However, these proteins were not precipitated when the complexes were denatured and reduced before immunoprecipitation. Using 1-ethyl 3-(3-dimethylaminopropyl)carbodiimide, we observed erythropoietin cross-linking with a protein of 66 kDa in addition to the 100- and 85-kDa proteins. Only the 66-kDa erythropoietin-cross-linked protein was immunoprecipitated by anti-receptor antibodies after denaturation and reduction of the complex. Thus, our results suggest that the 85- and 100-kDa proteins previously evidenced by cross-linking are associated with the cloned chain of the receptor to form a multimeric complex but these proteins seem immunologically unrelated to the cloned chain. We observed that reducing the length of molecules able to cross-link amino groups decreased the efficiency of cross-linking with the 100-kDa protein and only the 85-kDa protein was cross-linked with erythropoietin using 1,5-difluoro-2,4-dinitrobenzene. These results suggest that the 85- and 100-kDa proteins occupate slightly different positions relative to the erythropoietin molecule bound to the receptor.