Structural and functional characterisation of a novel peptide from the Australian sea anemone Actinia tenebrosa

Khaled A. Elnahriry, Dorothy C.C. Wai, Bankala Krishnarjuna, Noha N. Badawy, Balasubramanyam Chittoor, Christopher A. MacRaild, Billy J. Williams-Noonan, Joachim M. Surm, David K. Chalmers, Alan H. Zhang, Steve Peigneur, Mehdi Mobli, Jan Tytgat, Peter Prentis, Raymond S. Norton

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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 1H 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.

Original languageEnglish
Pages (from-to)104-112
Number of pages9
Publication statusPublished - 1 Oct 2019


  • Cysteine-containing peptide
  • Lipid interactions
  • NMR spectroscopy
  • Sea anemone
  • Structure

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