Department of Chemistry, Graduate School of Science, the University of Tokyo, 7-3-1 Bunkyo-ku, Tokyo 113-0033, Japan.
Molecules. 2013 Aug 30;18(9):10514-30. doi: 10.3390/molecules180910514.
The random non-standard peptide integrated discovery (RaPID) system has proven to be a powerful approach to discover de novo natural product-like macrocyclic peptides that inhibit protein functions. We have recently reported three macrocyclic peptides that bind to Pyrococcus furiosus multidrug and toxic compound extrusion (PfMATE) transporter and inhibit the transport function. Moreover, these macrocyclic peptides were successfully employed as cocrystallization ligands of selenomethionine-labeled PfMATE. In this report, we disclose the details of the RaPID selection strategy that led to the identification of these three macrocyclic peptides as well as a fourth macrocyclic peptide, MaD8, which is exclusively discussed in this article. MaD8 was found to bind within the cleft of PfMATE's extracellular side and blocked the path of organic small molecules being extruded. The results of an ethidium bromide efflux assay confirmed the efflux inhibitory activity of MaD8, whose behavior was similar to that of previously reported MaD5.
随机非标准肽综合发现（RaPID）系统已被证明是一种强大的方法，可以发现抑制蛋白质功能的新型天然产物样大环肽。我们最近报道了三种与 Pyrococcus furiosus 多药和毒性化合物外排（PfMATE）转运蛋白结合并抑制其转运功能的大环肽。此外，这些大环肽已成功用作硒代蛋氨酸标记的 PfMATE 的共结晶配体。在本报告中，我们将披露导致鉴定这三种大环肽以及第四种大环肽 MaD8 的 RaPID 选择策略的详细信息，MaD8 仅在本文中进行讨论。发现 MaD8 结合在 PfMATE 细胞外表面的裂缝内，并阻止有机小分子被排出。溴化乙锭外排测定的结果证实了 MaD8 的外排抑制活性，其行为与先前报道的 MaD5 相似。
