RGD-INDEPENDENT CELL-ADHESION TO THE CARBOXY-TERMINAL HEPARIN-BINDING FRAGMENT OF FIBRONECTIN INVOLVES HEPARIN-DEPENDENT AND HEPARIN-INDEPENDENT ACTIVITIES
RGD-INDEPENDENT CELL-ADHESION TO THE CARBOXY-TERMINAL HEPARIN-BINDING FRAGMENT OF FIBRONECTIN INVOLVES HEPARIN-DEPENDENT AND HEPARIN-INDEPENDENT ACTIVITIES
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DOI：
10.1083/jcb.110.3.777
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发表时间：
1990-03-01
影响因子：
7.8
通讯作者：
FURCHT, LT
中科院分区：
文献类型：
--
作者：
MCCARTHY, JB;SKUBITZ, APN;FURCHT, LT
细胞与细胞外基质组分如纤连蛋白的粘附具有复杂的基础，涉及与离散细胞表面大分子反应的分子上的多个决定簇。我们以前的研究结果表明，正常和转化细胞粘附和传播的33-kD肝素结合片段，源自特定亚型（A-链）的人纤连蛋白的羧基末端。该片段促进黑色素瘤的粘附和扩散，在一个乙酰-甘氨酰-乙酰-丝氨酸（RGDS）的独立方式，表明细胞粘附到纤连蛋白的这个区域是独立的典型的RGD/整合素介导的结合。最近鉴定了来自纤连蛋白该区域的两种合成肽，其在固相测定中结合[3 H]肝素并促进黑素瘤细胞的粘附和扩散（McCarthy，J. B.，M. K. Chelberg，D. J. Mickelson和L. T.弗赫特1988.生物化学。27：1380-1388）。目前的研究进一步确定了这些合成肽之一的细胞粘附和肝素结合特性。该肽称为肽I，具有序列YEKPGSP-PREVVPRPGV，代表人血浆纤连蛋白的残基1906-1924。除了以浓度依赖性方式促进不依赖于RGD的黑色素瘤粘附和扩散外，该肽还显著抑制细胞与33-kD片段或完整纤连蛋白的粘附。针对肽I产生的多克隆抗体也显着抑制细胞粘附的肽，33-kD片段，但黑色素瘤粘附纤连蛋白的影响很小。抗肽I抗体也部分抑制[3 H]肝素与纤连蛋白的结合，表明肽I代表完整分子上的主要肝素结合结构域。来自33-kD片段的另一种肽，称为CS 1（Humphries，M. J.，A.小森屋，S. K.秋山K.奥登和K. M.山田1987. J. Biol. Chem.，262：6886-6892）与肽I进行对比。而这两种肽促进RGD-非依赖性细胞粘附，肽CS 1未能结合肝素，和外源性肽CS 1未能抑制肽I介导的细胞粘附。结果表明，不同的肝素依赖性和非依赖性细胞粘附决定簇的33 kD片段，这两个都不相关的RGD依赖性整合素与纤连蛋白的相互作用的作用。
Cell adhesion to extracellular matrix components such as fibronectin has a complex basis, involving multiple determinants on the molecular that react with discrete cell surface macromolecules. Our previous results have demonstrated that normal and transformed cells adhere and spread on a 33-kD heparin binding fragment that originates from the carboxyterminal end of particular isoforms (A-chains) of human fibronectin. This fragment promotes melanoma adhesion and spreading in an arginyl-glycyl-aspartyl-serine (RGDS) independent manner, suggesting that cell adhesion to this region of fibronectin is independent of the typical RGD/integrin-mediated binding. Two synthetic peptides from this region of fibronectin were recently identified that bound [3H]heparin in a solid-phase assay and promoted the adhesion and spreading of melanoma cells (McCarthy, J. B., M. K. Chelberg, D. J. Mickelson, and L. T. Furcht. 1988. Biochemistry. 27:1380-1388). The current studies further define the cell adhesion and heparin binding properties of one of these synthetic peptides. This peptide, termed peptide I, has the sequence YEKPGSP-PREVVPRPRPGV and represents residues 1906-1924 of human plasma fibronectin. In addition to promoting RGD-independent melanoma adhesion and spreading in a concentration-dependent manner, this peptide significantly inhibited cell adhesion to the 33-kD fragment or intact fibronectin. Polyclonal antibodies generated against peptide I also significantly inhibited cell adhesion to the peptide, to the 33-kD fragment, but had minimal effect on melanoma adhesion to fibronectin. Anti-peptide I antibodies also partially inhibited [3H]heparin binding to fibronectin, suggesting that peptide I represents a major heparin binding domain on the intact molecule. The cell adhesion activity of another peptide from the 33-kD fragment, termed CS1 (Humphries, M. J., A. Komoriya, S. K. Akiyama, K. Olden, and K. M. Yamada. 1987. J. Biol. Chem., 262:6886-6892) was contrasted with peptide I. Whereas both peptides promoted RGD-independent cell adhesion, peptide CS1 failed to bind heparin, and exogenous peptide CS1 failed to inhibit peptide I-mediated cell adhesion. The results demonstrate a role for distinct heparin-dependent and -independent cell adhesion determinants on the 33-kD fragment, neither of which are related to the RGD-dependent integrin interaction with fibronectin.