EXPRESSION CLONING OF A FUNCTIONAL GLYCOPROTEIN LIGAND FOR P-SELECTIN
EXPRESSION CLONING OF A FUNCTIONAL GLYCOPROTEIN LIGAND FOR P-SELECTIN
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DOI：
10.1016/0092-8674(93)90327-m
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发表时间：
1993-12-17
期刊：
影响因子：
64.5
通讯作者：
LARSEN, GR
中科院分区：
文献类型：
--
作者：
SAKO, D;CHANG, XJ;LARSEN, GR
循环白细胞和内皮细胞之间最初的粘附相互作用部分由P-选择素介导。我们现在报告从HL-60 cDNA文库中表达克隆P-选择素的功能性配体。预测的氨基酸序列揭示了一种新的粘蛋白样跨膜蛋白。转染COS细胞与P-选择素的显著结合需要蛋白质配体和岩藻糖基转移酶的共表达。这种结合是钙依赖性的，可以被P-选择素的中和单克隆抗体抑制。共转染的COS细胞表达220 kd的同源二聚体的配体。可溶性配体构建体在COS细胞中与岩藻糖基转移酶共表达时，也介导P-选择素结合，并与特异性结合P-选择素的主要HL-60糖蛋白发生免疫交叉反应。
The initial adhesive interactions between circulating leukocytes and endothelia are mediated, in part, by P-selectin. We now report the expression cloning of a functional ligand for P-selectin from an HL-60 cDNA library. The predicted amino acid sequence reveals a novel mucin-like transmembrane protein. Significant binding of transfected COS cells to P-selectin requires coexpression of both the protein ligand and a fucosyltransferase. This binding is calcium dependent and can be inhibited by a neutralizing monoclonal antibody to P-selectin. Cotransfected COS cells express the ligand as a homodimer of 220 kd. A soluble ligand construct, when coexpressed with fucosyltransferase in COS cells, also mediates P-selectin binding and is immunocrossreactive with the major HL-60 glycoprotein that specifically binds P-selectin.