Identification of the receptor for erythropoietin by cross-linking to Friend virus-infected erythroid cells.
Identification of the receptor for erythropoietin by cross-linking to Friend virus-infected erythroid cells.
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通过与弗兰德病毒感染的红细胞交联来鉴定促红细胞生成素受体。
DOI：
10.1073/pnas.84.11.3690
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发表时间：
1987
影响因子：
11.1
通讯作者：
Luna,J
中科院分区：
文献类型：
--
作者：
Sawyer,ST;Krantz,SB;Luna,J
促红细胞生成素（Epo）是一种糖蛋白激素，其调节红系细胞发育并与发育中的红系细胞上的表面受体相互作用。在该实验室中，已经开发出具有对Epo有反应的相对纯的红系细胞群体的细胞系统。未成熟红系细胞获自感染Friend病毒贫血株的小鼠脾脏。本文研究了~（125）I标记的Epo（~（125）I-Epo）与这些细胞质膜的结合。在37 ℃下，125 I-Epo结合在20分钟内达到平衡。20%的受体与~（125）I-Epo结合的Kd为0.08 × 10 ~（-9）M，其余受体与~（125）I-Epo结合的Kd为0.6 × 10 ~（-9）M。在这项研究中，Epo的受体被确定为通过交联125 I-Epo的受体在完整的细胞和质膜制备使用二琥珀酰亚胺辛二酸酯。聚丙烯酰胺凝胶电泳显示100和85 kDa的两个标记带。85-kDa条带的标记程度（65%）高于100-kDa条带。当结合混合物中含有增加量的未标记Epo时，这两条带同样减少，表明125 I-Epo与受体的特异性相互作用。
Erythropoietin (Epo) is a glycoprotein hormone that regulates erythroid development and interacts with surface receptors on developing erythroid cells. In this laboratory, a cell system with a relatively pure population of erythroid cells that respond to Epo has been developed. Immature erythroid cells are obtained from the spleens of mice infected with the anemia strain of Friend virus. The binding of 125I-labeled Epo (125I-Epo) to plasma membranes from these cells was studied in this investigation. 125I-Epo binding reached equilibrium within 20 min at 37 degrees C. Twenty percent of the receptors bound 125I-Epo with a Kd of 0.08 X 10(-9) M, while the remaining receptors bound the hormone with a Kd of 0.6 X 10(-9) M. In this study, a receptor for Epo was identified by cross-linking 125I-Epo to the receptor in intact cells and plasma membrane preparations using disuccinimidyl suberate. Polyacrylamide gel electrophoresis revealed two labeled bands of 100 and 85 kDa. The 85-kDa band was more heavily labeled (65%) than the 100-kDa band. Both bands were equally decreased when increasing amounts of unlabeled Epo were included in the binding mixture, indicating a specific interaction of 125I-Epo with the receptor.