Recombinant Collagen Engineered to Bind to Discoidin Domain Receptor Functions as a Receptor Inhibitor.
Recombinant Collagen Engineered to Bind to Discoidin Domain Receptor Functions as a Receptor Inhibitor.
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DOI：
10.1074/jbc.m115.674507
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发表时间：
2016-02-26
期刊：
影响因子：
--
通讯作者：
Brodsky B
中科院分区：
文献类型：
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作者：
An B;Abbonante V;Xu H;Gavriilidou D;Yoshizumi A;Bihan D;Farndale RW;Kaplan DL;Balduini A;Leitinger B;Brodsky B
细菌胶原样蛋白Scl 2已被开发为重组胶原模型系统，以宿主人胶原配体结合序列，目的是产生具有选择性胶原生物活性的生物材料。整合素、纤连蛋白、肝素和MMP-1在人胶原中的定义的结合位点已被引入细菌胶原的三螺旋结构域，并导致预期的生物活性。活性的模块化插入在此延伸至盘状结构域受体（DDRs），其是胶原蛋白活化的受体酪氨酸激酶。将来自人胶原蛋白III的DDR结合序列插入细菌胶原蛋白中导致特异性受体结合。然而，即使在最高的可测试浓度下，该构建体也无法刺激DDR自磷酸化。在大肠杆菌中表达的重组胶原不含羟脯氨酸（Hyp），并且互补合成肽的研究表明，在关键的Gly-Val-Met-Gly-Phe-Hyp位置用Pro替换Hyp降低了DDR结合亲和力，因此需要更高的浓度用于诱导受体活化。重组细菌胶原蛋白结合DDRs而不诱导激酶活化的能力表明其可以干扰动物胶原蛋白和DDRs之间的相互作用，并且这种抑制作用在体外和细胞迁移测定中得到证实。这项研究表明，重组胶原蛋白可以补充合成肽的构效关系的研究，这个系统有可能引入或抑制特定的生物活性。
A bacterial collagen-like protein Scl2 has been developed as a recombinant collagen model system to host human collagen ligand-binding sequences, with the goal of generating biomaterials with selective collagen bioactivities. Defined binding sites in human collagen for integrins, fibronectin, heparin, and MMP-1 have been introduced into the triple-helical domain of the bacterial collagen and led to the expected biological activities. The modular insertion of activities is extended here to the discoidin domain receptors (DDRs), which are collagen-activated receptor tyrosine kinases. Insertion of the DDR-binding sequence from human collagen III into bacterial collagen led to specific receptor binding. However, even at the highest testable concentrations, the construct was unable to stimulate DDR autophosphorylation. The recombinant collagen expressed in Escherichia coli does not contain hydroxyproline (Hyp), and complementary synthetic peptide studies showed that replacement of Hyp by Pro at the critical Gly-Val-Met-Gly-Phe-Hyp position decreased the DDR-binding affinity and consequently required a higher concentration for the induction of receptor activation. The ability of the recombinant bacterial collagen to bind the DDRs without inducing kinase activation suggested it could interfere with the interactions between animal collagen and the DDRs, and such an inhibitory role was confirmed in vitro and with a cell migration assay. This study illustrates that recombinant collagen can complement synthetic peptides in investigating structure-activity relationships, and this system has the potential for the introduction or inhibition of specific biological activities.