Enzyme-Instructed Self-Assembly of Small D-Peptides as a Multiple-Step Process for Selectively Killing Cancer Cells.
Enzyme-Instructed Self-Assembly of Small D-Peptides as a Multiple-Step Process for Selectively Killing Cancer Cells.
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DOI：
10.1021/jacs.5b13541
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发表时间：
2016-03-23
影响因子：
15
通讯作者：
Xu B
中科院分区：
文献类型：
--
作者：
Zhou J;Du X;Yamagata N;Xu B
选择性抑制癌细胞仍然是化疗中的一个挑战。在这里，我们报告了酶指导的自组装(EISA)的分子和细胞验证，作为一种多步骤过程，选择性地杀死过表达碱性磷酸酶(ALP)的癌细胞。我们设计并合成了两种含有一个或两个磷酸酪氨酸残基，N-末端带有一个萘基的d-四肽。在酶去磷酸化后，这些d-四肽在水中变成自组装分子，形成纳米纤维。将这些d-四肽与几个癌细胞株和一个正常细胞系孵育，未磷酸化的d-四肽对所有的细胞株都是无害的，单磷酸化和二磷酸化的d-四肽选择性地抑制癌细胞，而不是正常细胞。单磷酸化的d-四肽比二磷酸化的d-四肽显示出更强的抑制活性；表达更高水平的ALP的癌细胞更容易被磷酸化的d-四肽抑制；具有更高自组装能力的d-四肽的前体显示出更高的抑制活性。这些结果证实了酶反应和自组装的重要作用。利用Alps的非竞争性抑制剂和荧光d-四肽，我们描述了酶催化的去磷酸化和自组装步骤，共同导致了d-四肽纳米纤维的定位，从而杀死了癌细胞。我们发现，细胞死亡模式可能与细胞类型有关，并证明了纳米纤维与死亡受体之间的相互作用。这项工作展示了一种范式转换和仿生方法，并为开发时空定义的超分子过程/组装作为潜在的抗癌治疗药物提供了有用的分子见解。
Selective inhibition of cancer cells remains a challenge in chemotherapy. Here we report the molecular and cellular validation of enzyme-instructed self-assembly (EISA) as a multiple step process for selectively killing cancer cells that overexpress alkaline phosphatases (ALPs). We design and synthesize two kinds of d-tetrapeptide containing one or two phosphotyrosine residues and with the N-terminal capped by a naphthyl group. Upon enzymatic dephosphorylation, these d-tetrapeptides turn into self-assembling molecules to form nanofibers in water. Incubating these d-tetrapeptides with several cancer cell lines and one normal cell line, the unphosphorylated d-tetrapeptides are innocuous to all the cell lines, the mono- and diphosphorylated d-tetrapeptides selectively inhibit the cancer cells, but not the normal cell. The monophosphorylated d-tetrapeptides exhibit more potent inhibitory activity than the diphosphorylated d-tetrapeptides do; the cancer cell lines express higher level of ALPs are more susceptible to inhibition by the phosphorylated d-tetrapeptides; the precursors of d-tetrapeptides that possess higher self-assembling abilities exhibit higher inhibitory activities. These results confirm the important role of enzymatic reaction and self-assembly. Using uncompetitive inhibitors of ALPs and fluorescent d-tetrapeptides, we delineate that the enzyme catalyzed dephosphorylation and the self-assembly steps, together, result in the localization of the nanofibers of d-tetrapeptides for killing the cancer cells. We find that the cell death modality likely associates with the cell type and prove the interactions between nanofibers and the death receptors. This work illustrates a paradigm-shifting and biomimetic approach and contributes useful molecular insights for the development of spatiotemporal defined supramolecular processes/assemblies as potential anticancer therapeutics.