Isolation of two insecticidal toxins from venom of the Australian theraphosid spider Coremiocnemis tropix

Maria P. Ikonomopoulou, Jennifer J. Smith, Volker Herzig, Sandy S. Pineda, Sławomir Dziemborowicz, Sing Yan Er, Thomas Durek, John Gilchrist, Paul F. Alewood, Graham M. Nicholson, Frank Bosmans, Glenn F. King

Research output: Contribution to journalArticle

Abstract

Sheep flystrike is caused by parasitic flies laying eggs on soiled wool or open wounds, after which the hatched maggots feed on the sheep flesh and often cause large lesions. It is a significant economic problem for the livestock industry as infestations are difficult to control due to ongoing cycles of larval development into flies followed by further egg laying. We therefore screened venom fractions from the Australian theraphosid spider Coremiocnemis tropix to identify toxins active against the sheep blowfly Lucilia cuprina, which is the primary cause of flystrike in Australia. This screen led to isolation of two insecticidal peptides, Ct1a and Ct1b, that are lethal to blowflies within 24 h of injection. The primary structure of these peptides was determined using a combination of Edman degradation and sequencing of a C. tropix venom-gland transcriptome. Ct1a and Ct1b contain 39 and 38 amino acid residues, respectively, including six cysteine residues that form three disulfide bonds. Recombinant production in bacteria (Escherichia coli) resulted in low yields of Ct1a whereas solid-phase peptide synthesis using native chemical ligation produced sufficient quantities of Ct1a for functional analyses. Synthetic Ct1a had no effect on voltage-gated sodium channels from the American cockroach Periplanata americana or the German cockroach Blattella germanica, but it was lethal to sheep blowflies with an LD50 of 1687 pmol/g.

Original languageEnglish (US)
Pages (from-to)62-70
Number of pages9
JournalToxicon
Volume123
DOIs
StatePublished - Dec 1 2016

Fingerprint

Spiders
Venoms
Sheep
Peptides
Diptera
Voltage-Gated Sodium Channels
Blattellidae
Periplaneta
Wool
Disulfides
Farms
Escherichia coli
Solid-Phase Synthesis Techniques
Cysteine
Lethal Dose 50
Bacteria
Livestock
Transcriptome
Eggs
Ovum

Keywords

  • Bioinsecticide
  • Coremiocnemis tropix
  • Flystrike
  • Insecticidal peptide
  • Lucilia cuprina
  • Spider venom

ASJC Scopus subject areas

  • Toxicology

Cite this

Ikonomopoulou, M. P., Smith, J. J., Herzig, V., Pineda, S. S., Dziemborowicz, S., Er, S. Y., ... King, G. F. (2016). Isolation of two insecticidal toxins from venom of the Australian theraphosid spider Coremiocnemis tropix. Toxicon, 123, 62-70. https://doi.org/10.1016/j.toxicon.2016.10.013

Isolation of two insecticidal toxins from venom of the Australian theraphosid spider Coremiocnemis tropix. / Ikonomopoulou, Maria P.; Smith, Jennifer J.; Herzig, Volker; Pineda, Sandy S.; Dziemborowicz, Sławomir; Er, Sing Yan; Durek, Thomas; Gilchrist, John; Alewood, Paul F.; Nicholson, Graham M.; Bosmans, Frank; King, Glenn F.

In: Toxicon, Vol. 123, 01.12.2016, p. 62-70.

Research output: Contribution to journalArticle

Ikonomopoulou, MP, Smith, JJ, Herzig, V, Pineda, SS, Dziemborowicz, S, Er, SY, Durek, T, Gilchrist, J, Alewood, PF, Nicholson, GM, Bosmans, F & King, GF 2016, 'Isolation of two insecticidal toxins from venom of the Australian theraphosid spider Coremiocnemis tropix', Toxicon, vol. 123, pp. 62-70. https://doi.org/10.1016/j.toxicon.2016.10.013
Ikonomopoulou MP, Smith JJ, Herzig V, Pineda SS, Dziemborowicz S, Er SY et al. Isolation of two insecticidal toxins from venom of the Australian theraphosid spider Coremiocnemis tropix. Toxicon. 2016 Dec 1;123:62-70. https://doi.org/10.1016/j.toxicon.2016.10.013
Ikonomopoulou, Maria P. ; Smith, Jennifer J. ; Herzig, Volker ; Pineda, Sandy S. ; Dziemborowicz, Sławomir ; Er, Sing Yan ; Durek, Thomas ; Gilchrist, John ; Alewood, Paul F. ; Nicholson, Graham M. ; Bosmans, Frank ; King, Glenn F. / Isolation of two insecticidal toxins from venom of the Australian theraphosid spider Coremiocnemis tropix. In: Toxicon. 2016 ; Vol. 123. pp. 62-70.
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abstract = "Sheep flystrike is caused by parasitic flies laying eggs on soiled wool or open wounds, after which the hatched maggots feed on the sheep flesh and often cause large lesions. It is a significant economic problem for the livestock industry as infestations are difficult to control due to ongoing cycles of larval development into flies followed by further egg laying. We therefore screened venom fractions from the Australian theraphosid spider Coremiocnemis tropix to identify toxins active against the sheep blowfly Lucilia cuprina, which is the primary cause of flystrike in Australia. This screen led to isolation of two insecticidal peptides, Ct1a and Ct1b, that are lethal to blowflies within 24 h of injection. The primary structure of these peptides was determined using a combination of Edman degradation and sequencing of a C. tropix venom-gland transcriptome. Ct1a and Ct1b contain 39 and 38 amino acid residues, respectively, including six cysteine residues that form three disulfide bonds. Recombinant production in bacteria (Escherichia coli) resulted in low yields of Ct1a whereas solid-phase peptide synthesis using native chemical ligation produced sufficient quantities of Ct1a for functional analyses. Synthetic Ct1a had no effect on voltage-gated sodium channels from the American cockroach Periplanata americana or the German cockroach Blattella germanica, but it was lethal to sheep blowflies with an LD50 of 1687 pmol/g.",
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