A disulfide-stabilised helical hairpin fold in acrorhagin I: An emerging structural motif in peptide toxins

[Display omitted] •The first structure of a peptide from sea anemone acrorhagi is reported.•Acrorhagin I has a similar solution structure to that of B–IV, α2p8, WaTx and globins, but with minimal or no sequence homology.•Acrorhagin I is not active against KV1 channels and showed no growth inhibitory activity against bacteria and fungi.•Acrorhagin I binds divalent metal ions. Acrorhagin I (U-AITX-Aeq5a) is a disulfide–rich peptide identified in the aggressive organs (acrorhagi) of the sea anemone Actinia equina. Previous studies (Toxicon 2005, 46:768–74) found that the peptide is toxic in crabs, although the structural and functional properties of acrorhagin I have not been reported. In this work, an Escherichia coli (BL21 strain) expression system was established for the preparation of 13C,15N–labelled acrorhagin I, and the solution structure was determined using NMR spectroscopy. Structurally, acrorhagin I is similar to B–IV toxin from the marine worm Cerebratulus lacteus (PDB id 1VIB), with a well–defined helical hairpin structure stabilised by four intramolecular disulfide bonds. The recombinant peptide was tested in patch–clamp electrophysiology assays against voltage-gated potassium and sodium channels, and in bacterial and fungal growth inhibitory assays and haemolytic assays. Acrorhagin I was not active against any of the ion channels tested and showed no activity in functional assays, indicating that this peptide may possess a different biological function. Metal ion interaction studies using NMR spectroscopy showed that acrorhagin I bound zinc and nickel, suggesting that its function might be modulated by metal ions or that it may be involved in regulating metal ion levels and their transport. The similarity between the structure of acrorhagin I and that of B-IV toxin from a marine worm suggests that this fold may prove to be a recurring motif in disulfide-rich peptides from marine organisms.