Vascular endothelial cell adhesion and spreading promoted by the peptide REDV of the IIICS region of plasma fibronectin is mediated by integrin alpha 4 beta 1.
Vascular endothelial cell adhesion and spreading promoted by the peptide REDV of the IIICS region of plasma fibronectin is mediated by integrin alpha 4 beta 1.
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DOI：
10.1016/s0021-9258(19)49672-5
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发表时间：
1992-07
期刊：
影响因子：
--
通讯作者：
Stephen Massia;Jeffrey A. Hubbell
中科院分区：
文献类型：
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作者：
Stephen Massia;Jeffrey A. Hubbell
我们最近报道了人脐静脉内皮细胞（HUVEC）在含有共价接枝的Arg-Glu-Asp-Val（REDV）肽的底物上的附着和铺展（Hubbell，J.A.，Massia，S. P.，Desai，N. P.，和德拉姆海勒，P. D.（1991）Bio/Technology 9，568-572）。据报道，该肽是纤连蛋白可变剪接的III型连接片段（IIICS）区域的CS5位点内的最小活性序列，并且整合素α 4 β 1已被鉴定为黑素瘤细胞上该位点的受体。整联蛋白α 4 β 1也被鉴定为神经嵴起源细胞、黑素瘤细胞、淋巴细胞和造血干细胞上IIICS区域中CS 1位点的受体。在这项研究中，我们证明，这种整合素也作为受体对HUVEC的肽REDV从CS5网站。通过全细胞酶联免疫吸附测定，显示α 4亚基在HUVEC膜上表达。针对整联蛋白亚基α 4和β 1的抗功能抗体抑制细胞粘附在REDV移植的基底上，但不抑制在RGD移植的基底上。α 4亚基定位成纤维状结构内的扩展细胞的REDV接枝基板上，但不是在扩展细胞的RGD接枝基板上。通过REDV配体亲和层析从HUVEC提取物中分离出两种蛋白质（144和120 kDa），并通过免疫沉淀和Western印迹证明其为整合素亚基α 4（144 kDa）和β 1（120 kDa）;此外，免疫沉淀分析表明亚基形成复合物。HUVEC与REDV-接枝基质的结合被可溶性REDV和RGD抑制，表明粘附是生物特异性的，并且REDV肽是RGD样的。在这份报告中，我们证明了第一次，α 4是存在于内皮细胞膜，在以前的报告相反，由他人，和整合素α 4 β 1是受体的REDV介导的粘附到IIICS区域的血浆纤连蛋白区域。
We have recently reported the attachment and spreading of human umbilical vein endothelial cells (HUVECs) upon substrates containing covalently grafted Arg-Glu-Asp-Val (REDV) peptide (Hubbell, J. A., Massia, S. P., Desai, N. P., and Drumheller, P. D. (1991) Bio/Technology 9, 568-572). This peptide has been reported to be the minimal active sequence within the CS5 site of the alternatively spliced type III connecting segment (IIICS) region of fibronectin, and the integrin alpha 4 beta 1 has been identified as the receptor on melanoma cells for this site. The integrin alpha 4 beta 1 has also been identified as the receptor for the CS1 site in the IIICS region on cells of neural crest origin, melanoma cells, lymphocytes, and hematopoietic stem cells. In this study, we demonstrate that this integrin also serves as a receptor on HUVECs for the peptide REDV from the CS5 site. The alpha 4 subunit was shown to be expressed upon HUVEC membranes by whole-cell enzyme-linked immunosorbent assay. Antifunctional antibodies directed against integrin subunits alpha 4 and beta 1 inhibited cell adhesion on REDV-grafted substrates, but not on RGD-grafted substrates. The alpha 4 subunit localized into fibrillar structures within spread cells on the REDV-grafted substrates, but not within spread cells on RGD-grafted substrates. Two proteins (144 and 120 kDa) were isolated from HUVEC extracts by REDV ligand affinity chromatography and were demonstrated by immunoprecipitation and Western blot to be the integrin subunits alpha 4 (144 kDa) and beta 1 (120 kDa); furthermore, the immunoprecipitation analyses demonstrated that the subunits formed a complex. HUVEC binding to REDV-grafted substrates was inhibited by both soluble REDV and RGD, demonstrating that adhesion was biospecific and that the REDV peptide is RGD-like. In this report we demonstrate for the first time that alpha 4 is present in the endothelial cell membrane, in contrast to previous reports by others, and that integrin alpha 4 beta 1 is the receptor for REDV-mediated adhesion to the IIICS region of region of plasma fibronectin.