Identification of a multifunctional, cell-binding peptide sequence from the a1(NC1) of type IV collagen.
Identification of a multifunctional, cell-binding peptide sequence from the a1(NC1) of type IV collagen.
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DOI：
10.1083/jcb.111.4.1583
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发表时间：
1990-10
影响因子：
7.8
通讯作者：
Furcht, L T
中科院分区：
文献类型：
--
作者：
Tsilibary, E C;Reger, L A;Vogel, A M;Koliakos, G G;Anderson, S S;Charonis, A S;Alegre, J N;Furcht, L T
我们之前已经从 IV 型胶原蛋白中鉴定出三种独特的氨基酸序列，它们特异性地与肝素结合，并且还抑制肝素与完整的 IV 型胶原蛋白的结合。这些化学合成结构域之一，肽 Hep-I，具有序列 TAGSCLRKFSTM，源自 IV 型胶原蛋白的 a1（非胶原 [NC1]）链（Koliakos, G. G., K. K. Koliakos, L. T. Furcht, L. A. Reger, and E. C. Tsilibary. 1989. J. Biol. Chem. 264：2313-2323）。我们在本报告中描述，在固相测定中，这种相同的肽也以剂量依赖性和特异性的方式与完整的 IV 型胶原蛋白结合。肽 Hep-I 和 IV 型胶原蛋白在溶液中的相互作用导致这种基底膜糖蛋白的组装过程受到抑制。因此，当这种蛋白质聚合形成网络时，肽 Hep-I 应该代表 IV 型胶原蛋白中的主要识别位点。此外，固相固定肽Hep-I能够促进牛主动脉内皮细胞的粘附和铺展。当存在于溶液中时，肽 Hep-I 以剂量依赖性方式竞争这些细胞与 IV 型胶原和 NC1 结构域包被的基质的结合。此外，溶液中的放射性标记肽Hep-I也以剂量依赖性和特异性方式与内皮细胞结合。过量的未标记肽可以抑制放射性标记的 Hep-I 与内皮细胞的结合。最后，在肝素或软骨素/硫酸皮肤素糖胺聚糖侧链存在的情况下，内皮细胞与肽Hep-I和NC1结构域包被的底物的结合也受到抑制。我们得出结论，肽 Hep-I 应该具有多种功能。这种 IV 型胶原衍生序列在自缔合、肝素结合以及细胞结合和粘附等多种现象中的作用，使得 Hep-I 成为参与确定基底膜超微结构以及细胞与含 IV 型胶原基质的相互作用的关键域。
We have previously identified three distinctive amino acid sequences from type IV collagen which specifically bound to heparin and also inhibited the binding of heparin to intact type IV collagen. One of these chemically synthesized domains, peptide Hep-I, has the sequence TAGSCLRKFSTM and originates from the a1(noncollagenous [NC1]) chain of type IV collagen (Koliakos, G. G., K. K. Koliakos, L. T. Furcht, L. A. Reger, and E. C. Tsilibary. 1989. J. Biol. Chem. 264:2313-2323). We describe in this report that this same peptide also bound to intact type IV collagen in solid-phase assays, in a dose-dependent and specific manner. Interactions between peptide Hep-I and type IV collagen in solution resulted in inhibition of the assembly process of this basement membrane glycoprotein. Therefore, peptide Hep-I should represent a major recognition site in type IV collagen when this protein polymerizes to form a network. In addition, solid phase- immobilized peptide Hep-I was able to promote the adhesion and spreading of bovine aortic endothelial cells. When present in solution, peptide Hep-I competed for the binding of these cells to type IV collagen- and NC1 domain-coated substrata in a dose-dependent manner. Furthermore, radiolabeled peptide Hep-I in solution also bound to endothelial cells in a dose-dependent and specific manner. The binding of radiolabeled Hep-I to endothelial cells could be inhibited by an excess of unlabeled peptide. Finally, in the presence of heparin or chondroitin/dermatan sulfate glycosaminoglycan side chains, the binding of endothelial cells to peptide Hep-I and NC1 domain-coated substrates was also inhibited. We conclude that peptide Hep-I should have a number of functions. The role of this type IV collagen-derived sequence in such diverse phenomena as self-association, heparin binding and cell binding and adhesion makes Hep-I a crucial domain involved in the determination of basement membrane ultrastructure and cellular interactions with type IV collagen-containing matrices.