Identification of a novel dual E- and N-cadherin antagonist
Identification of a novel dual E- and N-cadherin antagonist
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DOI：
10.1016/j.peptides.2009.05.010
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发表时间：
2009-08-01
期刊：
影响因子：
3
通讯作者：
Blaschuk, Orest W.
中科院分区：
文献类型：
--
作者：
Devemy, Emmanuelle;Blaschuk, Orest W.
E-钙粘附素和N-钙粘附素是与钙离子相关的细胞黏附分子，对多种生物学和疾病过程产生影响。因此，这些钙粘附素的拮抗剂可以被设想为治疗有用的药物。我们已经利用噬菌体展示技术发现了这样的拮抗剂。以人E-钙粘蛋白胞外区与人免疫球蛋白G1Fc段(E-cad/Fc)的融合蛋白为抗原，构建了抗人E-cad/Fc嵌合蛋白的噬菌体抗体库。所有被分离的噬菌体克隆还结合了一个由人N-钙粘蛋白胞外区与人免疫球蛋白G1 Fc片段(N-cad/Fc)融合而成的嵌合蛋白。用H-SWELYYPLRANL-NH2合成噬菌体克隆展示的多肽，经表面等离子体共振谱测定，E-和N-cad/Fc嵌合蛋白的亲和力(K-D)分别为9.4um M和323 nM。该多肽还能阻断E-和N-cad/Fc嵌合蛋白包裹的微球的聚集，以及对MCF-7和MDA-MB435人乳腺癌细胞的聚集(这些细胞分别表达E-和N-钙粘素)。最后，我们发现该肽破坏了MCF-7和MDA-MB435细胞的单层。证明该多肽H-SWELYPLRANL-NH2是一种具有生物活性的E-和N-钙粘附素双重拮抗剂。这种拮抗剂在各种生物学环境中都有应用。(C)2009 Elsevier Inc.保留所有权利。
E- and N-cadherin are related calcium-dependent cell adhesion molecules that exert an influence over multiple biological and disease processes. Antagonists of these cadherins can therefore be envisaged as therapeutically useful drugs. We have used phage display technology to discover such antagonists. A peptide phage library was screened against a chimeric protein composed of the human E-cadherin ectodomain fused to the Fc fragment of human immunoglobulin G1 (E-cad/Fc). All of the phage clones that were isolated also bound a chimeric protein composed of the human N-cadherin ectodomain fused to the Fc fragment of human immunoglobulin G1 (N-cad/Fc). A peptide displayed by several of the isolated phage clones was synthesized (H-SWELYYPLRANL-NH2) and found to bind both E- and N-cad/Fc chimeric proteins with affinities (K-D) of 9.4 mu M and 323 nM, respectively, as judged by surface plasmon resonance spectroscopy. This peptide was also capable of blocking the aggregation of E- and N-cad/Fc chimeric protein-coated beads, as well as the aggregation of MCF-7 and MDA-MB435 human breast cancer cells (these cells express E- and N-cadherin, respectively). Finally, we showed that the peptide disrupted MCF-7 and MDA-MB435 cell monolayers. The peptide, H-SWELYYPLRANL-NH2 thus proved to be a biologically active, dual E- and N-cadherin antagonist. Such an antagonist has application in a wide variety of biological contexts. (C) 2009 Elsevier Inc. All rights reserved.