A novel synthetic peptide from the B1 chain of laminin with heparin-binding and cell adhesion-promoting activities.
A novel synthetic peptide from the B1 chain of laminin with heparin-binding and cell adhesion-promoting activities.
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来自层粘连蛋白的B1链的一种新型合成肽，具有肝素结合和细胞粘附活性。
DOI：
10.1083/jcb.107.3.1253
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发表时间：
1988-09
影响因子：
7.8
通讯作者：
Furcht, L T
中科院分区：
文献类型：
--
作者：
Charonis, A S;Skubitz, A P;Koliakos, G G;Reger, L A;Dege, J;Vogel, A M;Wohlhueter, R;Furcht, L T
最近使用固相结合测定和电子显微镜的研究表明，在侧短臂的内球和层粘连蛋白的交叉区域之间存在肝素结合域。利用层粘连蛋白B1链的氨基酸序列信息，从亲水指数低、赖氨酸和/或精氨酸密度高的区域合成了几种多肽。其中一种肽F-9 （RYVVLPRPVCFEKGMNYTVR）来源于外侧短臂的内球状结构域，显示出与肝素的特异性结合。这在直接固相结合试验中进行了测试，通过将肽涂在硝化纤维素或聚苯乙烯上，并在间接竞争试验中进行了测试，其中肽在溶液中，层粘连蛋白或肝素固定在固体载体上。当在竞争试验中使用肝素而不是除肝素（硫酸葡聚糖、硫酸皮聚糖）以外的其他糖胺聚糖时，[3H]肝素与肽F-9的结合显著降低。通过乙酰化修饰肽F-9的游离氨基，取消了其抑制聚苯乙烯表面上[3H]肝素与层粘连蛋白结合的能力。肽F-9促进各种细胞系（黑色素瘤、纤维肉瘤、胶质瘤、嗜铬细胞瘤）和主动脉内皮细胞的粘附。此外，当肽F-9存在于溶液中时，它可以抑制黑色素瘤细胞对层粘连蛋白涂层底物的粘附。这些发现表明肽F-9在层粘连蛋白上定义了一个新的肝素结合和细胞粘附促进位点。
Recent studies using solid-phase-binding assays and electron microscopy suggested the presence of a heparin-binding domain between the inner globule of a lateral short arm and the cross region of laminin. Using the information from the amino acid sequence of the B1 chain of laminin, several peptides were synthesized from areas with a low hydropathy index and a high density of lysines and/or arginines. One of these, peptide F-9 (RYVVLPRPVCFEKGMNYTVR), which is derived from the inner globular domain of the lateral short arm, demonstrated specific binding to heparin. This was tested in direct solid-phase binding assays by coating the peptide either on nitrocellulose or on polystyrene and in indirect competition assays where the peptide was in solution and either laminin or heparin was immobilized on a solid support. The binding of [3H]heparin to peptide F-9 was dramatically reduced when heparin but not other glycosaminoglycans other than heparin (dextran sulfate, dermatan sulfate) were used in competition assays. Modification of the free amino groups of peptide F-9 by acetylation abolished its ability to inhibit the binding of [3H]heparin to laminin on polystyrene surfaces. Peptide F-9 promoted the adhesion of various cell lines (melanoma, fibrosarcoma, glioma, pheochromocytoma) and of aortic endothelial cells. Furthermore, when peptide F-9 was present in solution, it inhibited the adhesion of melanoma cells to laminin-coated substrates. These findings suggest that peptide F-9 defines a novel heparin-binding and cell adhesion- promoting site on laminin.