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Expression of a soluble functional form of the integrin alpha(4)beta(1) in mammalian cells

机译:整合素α(4)beta(1)的可溶性功能形式在哺乳动物细胞中的表达

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The integrin alpha(4)beta(1)(VLA4) has been expressed as a soluble, active, heterodimeric immunoglobulin fusion protein, cDNAs encoding the extracellular domains of the human alpha(4) and beta(1) subunits were fused to the genomic DNA encoding the human yl immunoglobulin Fc domain and functional integrin fusion protein was expressed as a secreted, soluble molecule from a range of mammalian cell lines. Specific mutations were introduced into the Fc region of the molecules to promote alpha(4)beta(1) heterodimer formation. The soluble alpha(4)beta(1)-Fc fusion protein exhibited divalent cation dependent binding to VCAM-1, which was blocked by the appropriate function blocking antibodies. The apparent K-d for VCAM-1 binding were similar for both the soluble and native forms of a(4)beta(1). In addition, the integrin-Fc fusion was shown to stain cells expressing VCAM-1 on their surface by FACs analysis. This approach for expressing soluble alpha(4)beta(1) should be generally applicable to a range of integrins. [References: 44]
机译:整联蛋白alpha(4)beta(1)(VLA4)已表达为可溶性,活性,异二聚体免疫球蛋白融合蛋白,编码人alpha(4)和beta(1)亚基胞外域的cDNA与基因组融合编码人类yl免疫球蛋白Fc结构域和功能性整联蛋白融合蛋白的DNA被表达为一系列哺乳动物细胞系中分泌的可溶性分子。将特定的突变引入分子的Fc区,以促进alpha(4)beta(1)异二聚体的形成。可溶性α(4)β(1)-Fc融合蛋白表现出与VCAM-1的二价阳离子依赖性结合,该结合被适当的功能阻断抗体阻断。 VCAM-1结合的表观K-d对于a(4)beta(1)的可溶性和天然形式均相似。另外,整联蛋白-Fc融合物通过FAC分析显示对在其表面上表达VCAM-1的细胞染色。这种表达可溶性α(4)β(1)的方法通常应适用于一系列整联蛋白。 [参考:44]

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