CHARACTERIZATION OF A MEMBRANE-ASSOCIATED GLYCOPROTEIN COMPLEX IMPLICATED IN CELL-ADHESION TO FIBRONECTIN
CHARACTERIZATION OF A MEMBRANE-ASSOCIATED GLYCOPROTEIN COMPLEX IMPLICATED IN CELL-ADHESION TO FIBRONECTIN
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DOI：
10.1002/jcb.240280409
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发表时间：
1985-01-01
影响因子：
4
通讯作者：
YAMADA, KM
中科院分区：
文献类型：
--
作者：
HASEGAWA, T;HASEGAWA, E;YAMADA, KM
一个140 kDa的糖蛋白复合物纯化的单克隆抗体和牵连细胞粘附的细胞外分子纤连蛋白的特点。通过单克隆抗体JG 22 E纯化了三种主要的多肽组分，其表观分子量分别为155，000（条带1）、135，000（条带2）和120，000（条带3）。在二维凝胶电泳中，每个亚基在酸性等电点处以宽带或一系列斑点的形式迁移。用神经氨酸酶处理后，斑点集中在pH 6.2（条带1）、pH 5.6（条带2）和pH 5.3（条带3）附近。这3个主要条带通过一系列成对组合的二维肽图谱进行比较，发现是不同的蛋白质。在蔗糖梯度中，这些蛋白质作为复合物共迁移，在apprx处沉降。8.4 S，而分离的亚基在4.7-5.8 S处迁移。氨基酸分析显示，没有检测到的羟脯氨酸和组合物的特点是大量的半胱氨酸残基相比，平均蛋白质。结构上不同的糖蛋白的非共价复合物参与纤连蛋白与细胞的粘附相互作用。
A 140-kDa glycoprotein complex purified by a monoclonal antibody and implicated in cell adhesion to the extracellular molecule fibronectin was characterized. Three major polypeptide compoments were purified by monclonal antibody JG22E, which had apparent MW of 155,000 (band 1), 135,000 (band 2) and 120,000 (band 3). In 2-dimensional gel electrophoresis, each subunit migrated as either a broad band or a series of spots at acidic isoelectric points. After treatment with neuraminidase, the spots became focused around pH 6.2 (band 1), pH 5.6 (band 2) and pH 5.3 (band 3). These 3 major bands were compared by 2-dimensional peptide mapping in a series of pairwise combinations and were found to be distinct proteins. In sucrose gradients, these proteins co-migrated as a complex sedimenting at .apprx. 8.4 S either before or after affinity purification, whereas separated subunits migrated at 4.7-5.8 S. Amino acid analysis revealed no detectable hydroxyproline and a composition characterized by a substantial number of Cys residues compared to the average protein. A noncovalent complex of sturcutrally distinct glycoproteins is involved in adhesive interactions of fibronectin with cells.