IDENTIFICATION AND ISOLATION OF A 140-KD CELL-SURFACE GLYCOPROTEIN WITH PROPERTIES EXPECTED OF A FIBRONECTIN RECEPTOR
IDENTIFICATION AND ISOLATION OF A 140-KD CELL-SURFACE GLYCOPROTEIN WITH PROPERTIES EXPECTED OF A FIBRONECTIN RECEPTOR
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DOI：
10.1016/0092-8674(85)90322-8
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发表时间：
1985-01-01
期刊：
影响因子：
64.5
通讯作者：
RUOSLAHTI, E
中科院分区：
文献类型：
--
作者：
PYTELA, R;PIERSCHBACHER, MD;RUOSLAHTI, E
亲和层析用于鉴定纤连蛋白的假定细胞表面受体。使用大的细胞附着促进纤连蛋白片段作为亲和基质，并通过使用含有序列Arg-Gly-Asp的合成肽进行特异性洗脱，所述序列Arg-Gly-Asp衍生自纤连蛋白细胞附着位点中的细胞识别序列。从MG-63人骨肉瘤细胞的辛基葡糖苷提取物中提取的亲和基质结合了一个140 kd [千道尔顿]的蛋白质，并用合成肽Gly-Arg-Gly-Asp-Ser-Pro特异性洗脱。通过细胞表面特异性放射性碘标记140-kd蛋白，并以高效率掺入脂质体中。含有这种蛋白质的脂质体对纤连蛋白包被的表面表现出特异性的亲和力，这种结合可以被合成的细胞附着肽选择性地抑制，但不能被非活性肽抑制。麦胚凝集素-琼脂糖凝胶亲和层析表明，140-kd蛋白是一种糖蛋白，并结合纤连蛋白片段层析，得到高度富集的140-kd蛋白的制剂。这些特性表明，140 kd的糖蛋白是一种膜包埋的细胞表面蛋白直接参与细胞粘附到纤连蛋白底物的初始步骤。
Affinity chromatography was used to identify a putative cell surface receptor for fibronectin. A large cell-attachment-promoting fibronectin fragment was used as the affinity matrix, and specific elution was effected by using synthetic peptides containing the sequence Arg-Gly-Asp, which is derived from the cell recognition sequence in the fibronectin cell attachment site. A 140 kd [kilodalton] protein was bound by the affinity matrix from octylglucoside extracts of MG-63 human osteosarcoma cells and specifically eluted with the synthetic peptide Gly-Arg-Gly-Asp-Ser-Pro. The 140-kd protein was labeled by cell surface specific radioiodination and became incorporated into liposomes at a high efficiency. Liposomes containing this protein showed specific affinity toward fibronectin-coated surfaces, and this binding could be selectively inhibited by the synthetic cell-attachment peptide but not by inactive peptides. Affinity chromatography on wheat germ agglutinin-Sepharose showed that the 140-kd protein is a glycoprotein and, in combination with the fibronectin fragment chromatography, gave highly enriched preparations of the 140-kd protein. These properties suggest that the 140-kd glycoprotein is a membrane-embedded cell surface protein directly involved in the initial step of cell adhesion to fibronectin substrates.