Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences, Monash University, 381 Royal Parade, Parkville, VIC, 3052, Australia.
School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Australia; Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Australia.
Toxicon. 2019 Oct;168:104-112. doi: 10.1016/j.toxicon.2019.07.002. Epub 2019 Jul 11.
Sea anemone venoms have long been recognised as a rich source of peptides with interesting pharmacological and structural properties. Our recent transcriptomic studies of the Australian sea anemone Actinia tenebrosa have identified a novel 13-residue peptide, U-AITx-Ate1. U-AITx-Ate1 contains a single disulfide bridge and bears no significant homology to previously reported amino acid sequences of peptides from sea anemones or other species. We have produced U-AITx-Ate1 using solid-phase peptide synthesis, followed by oxidative folding and purification of the folded peptide using reversed-phase high-performance liquid chromatography. The solution structure of U-AITx-Ate1 was determined based on two-dimensional nuclear magnetic resonance spectroscopic data. Diffusion-ordered NMR spectroscopy revealed that U-AITx-Ate1 was monomeric in solution. Perturbations in the 1D H NMR spectrum of U-AITx-Ate1 in the presence of dodecylphosphocholine micelles together with molecular dynamics simulations indicated an interaction of U-AITx-Ate1 with lipid membranes, although no binding was detected to 100% POPC and 80% POPC: 20% POPG lipid nanodiscs by isothermal titration calorimetry. Functional assays were performed to explore the biological activity profile of U-AITx-Ate1. U-AITx-Ate1 showed no activity in voltage-clamp electrophysiology assays and no change in behaviour and mortality rates in crustacea. Moderate cytotoxic activity was observed against two breast cancer cell lines.
海葵毒液长期以来一直被认为是富含具有有趣药理学和结构特性的肽的丰富来源。我们最近对澳大利亚海葵 Actinia tenebrosa 的转录组研究鉴定了一种新型的 13 残基肽，U-AITx-Ate1。U-AITx-Ate1 含有一个单一的二硫键，与以前报道的来自海葵或其他物种的肽的氨基酸序列没有显著同源性。我们使用固相肽合成生产了 U-AITx-Ate1，然后使用反相高效液相色谱对折叠肽进行氧化折叠和纯化。根据二维核磁共振波谱数据确定了 U-AITx-Ate1 的溶液结构。扩散有序 NMR 光谱表明 U-AITx-Ate1 在溶液中为单体。U-AITx-Ate1 在十二烷基磷酸胆碱胶束存在下的 1D H NMR 光谱的扰动以及分子动力学模拟表明 U-AITx-Ate1 与脂质膜相互作用，尽管通过等温滴定量热法未检测到与 100%POPC 和 80%POPC:20%POPG 脂质纳米盘的结合。进行了功能测定以探索 U-AITx-Ate1 的生物学活性特征。U-AITx-Ate1 在电压钳电生理学测定中没有活性，在甲壳类动物中也没有行为和死亡率的变化。对两种乳腺癌细胞系表现出中等的细胞毒性活性。
