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

Research output: Contribution to journalArticleResearchpeer-review

Abstract

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
JournalToxicon
Volume168
DOIs
Publication statusPublished - 1 Oct 2019

Keywords

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

Cite this

Elnahriry, Khaled A. ; Wai, Dorothy C.C. ; Krishnarjuna, Bankala ; Badawy, Noha N. ; Chittoor, Balasubramanyam ; MacRaild, Christopher A. ; Williams-Noonan, Billy J. ; Surm, Joachim M. ; Chalmers, David K. ; Zhang, Alan H. ; Peigneur, Steve ; Mobli, Mehdi ; Tytgat, Jan ; Prentis, Peter ; Norton, Raymond S. / Structural and functional characterisation of a novel peptide from the Australian sea anemone Actinia tenebrosa. In: Toxicon. 2019 ; Vol. 168. pp. 104-112.
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abstract = "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.",
keywords = "Cysteine-containing peptide, Lipid interactions, NMR spectroscopy, Sea anemone, Structure",
author = "Elnahriry, {Khaled A.} and Wai, {Dorothy C.C.} and Bankala Krishnarjuna and Badawy, {Noha N.} and Balasubramanyam Chittoor and MacRaild, {Christopher A.} and Williams-Noonan, {Billy J.} and Surm, {Joachim M.} and Chalmers, {David K.} and Zhang, {Alan H.} and Steve Peigneur and Mehdi Mobli and Jan Tytgat and Peter Prentis and Norton, {Raymond S.}",
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Elnahriry, KA, Wai, DCC, Krishnarjuna, B, Badawy, NN, Chittoor, B, MacRaild, CA, Williams-Noonan, BJ, Surm, JM, Chalmers, DK, Zhang, AH, Peigneur, S, Mobli, M, Tytgat, J, Prentis, P & Norton, RS 2019, 'Structural and functional characterisation of a novel peptide from the Australian sea anemone Actinia tenebrosa' Toxicon, vol. 168, pp. 104-112. https://doi.org/10.1016/j.toxicon.2019.07.002

Structural and functional characterisation of a novel peptide from the Australian sea anemone Actinia tenebrosa. / Elnahriry, Khaled A.; Wai, Dorothy C.C.; Krishnarjuna, Bankala; Badawy, Noha N.; Chittoor, Balasubramanyam; MacRaild, Christopher A.; Williams-Noonan, Billy J.; Surm, Joachim M.; Chalmers, David K.; Zhang, Alan H.; Peigneur, Steve; Mobli, Mehdi; Tytgat, Jan; Prentis, Peter; Norton, Raymond S.

In: Toxicon, Vol. 168, 01.10.2019, p. 104-112.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Structural and functional characterisation of a novel peptide from the Australian sea anemone Actinia tenebrosa

AU - Elnahriry, Khaled A.

AU - Wai, Dorothy C.C.

AU - Krishnarjuna, Bankala

AU - Badawy, Noha N.

AU - Chittoor, Balasubramanyam

AU - MacRaild, Christopher A.

AU - Williams-Noonan, Billy J.

AU - Surm, Joachim M.

AU - Chalmers, David K.

AU - Zhang, Alan H.

AU - Peigneur, Steve

AU - Mobli, Mehdi

AU - Tytgat, Jan

AU - Prentis, Peter

AU - Norton, Raymond S.

PY - 2019/10/1

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N2 - 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.

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KW - Cysteine-containing peptide

KW - Lipid interactions

KW - NMR spectroscopy

KW - Sea anemone

KW - Structure

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U2 - 10.1016/j.toxicon.2019.07.002

DO - 10.1016/j.toxicon.2019.07.002

M3 - Article

VL - 168

SP - 104

EP - 112

JO - Toxicon : official journal of the International Society on Toxinology

JF - Toxicon : official journal of the International Society on Toxinology

SN - 0041-0101

ER -