Abstract
Spider venoms represent invaluable sources of biologically active compounds suitable for use in life science research and also having a significant potential for biotechnology and therapeutic applications. The methods reported herewith are based on our long experience of spider venom fractionation and peptides purification. We routinely screen new peptides for antimicrobial and insecticidal activities and our detailed protocols are also reported here. So far these have been tested on species of Central Asian and European spiders from the families Agelenidae, Eresidae, Gnaphosidae, Lycosidae, Miturgidae, Oxyopidae, Philodromidae, Pisauridae, Segestriidae, Theridiidae, Thomisidae, and Zodariidae. The reported protocols should be easily adaptable for use with other arthropod species.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Mintz, I.M., Venema, V.J., Swiderek, K.M., Lee, T.D., Bean, B.P., and Adams, M.E. (1992) P-type calcium channels blocked by the spider toxin omega-Aga-IVA. Nature 355, 827–829.
Suchyna, T.M., Johnson, J.H., Hamer, K. et al. (2000) Identification of a peptide toxin from Grammostola spatulata spider venom that blocks cation-selective stretch-activated channels. J. Gen. Physiol. 115, 583–598.
Estrada, G., Villegas, E., and Corzo, G. (2007) Spider venoms: a rich source of acylpolyamines and peptides as new leads for CNS drugs. Nat. Prod. Rep. 24, 145–161.
Mazzuca, M., Heurteaux, C., Alloui, A. et al. (2007) A tarantula peptide against pain via ASIC1a channels and opioid mechanisms. Nat. Neurosci. 10, 943–945.
Kuhn-Nentwig, L. (2003) Antimicrobial and cytolytic peptides of venomous arthropods. Cell. Mol. Life Sci. 60, 2651–2668.
Kozlov, S.A., Vassilevski, A.A., Feofanov, A.V., Surovoy, A.Y., Karpunin, D.V., and Grishin, E.V. (2006) Latarcins, antimicrobial and cytolytic peptides from the venom of the spider Lachesana tarabaevi (Zodariidae) that exemplify biomolecular diversity. J. Biol. Chem. 281, 20983–20992.
Nicholson, G.M. (2007) Insect-selective spider toxins targeting voltage-gated sodium channels. Toxicon 49, 490–512.
King, G.F. (2007) Modulation of insect Ca(v) channels by peptidic spider toxins. Toxicon 49, 513–530.
Sollod, B.L., Wilson, D., Zhaxybayeva, O., Gogarten, J.P., Drinkwater, R., and King, G.F. (2005) Were arachnids the first to use combinatorial peptide libraries? Peptides 26, 131–139.
Escoubas, P. and Rash, L. (2004) Tarantulas: eight-legged pharmacists and combinatorial chemists. Toxicon 43, 555–574.
Escoubas, P., Sollod, B., and King, G.F. (2006) Venom landscapes: mining the complexity of spider venoms via a combined cDNA and mass spectrometric approach. Toxicon 47, 650–663.
Vassilevski, A.A., Kozlov, S.A., Samsonova, O.V. et al. (2008) Cyto-insectotoxins, a novel class of cytolytic and insecticidal peptides from spider venom. Biochem. J. 411, 687–696.
Lipkin, A., Kozlov, S., Nosyreva, E., Blake, A., Windass, J.D., and Grishin, E. (2002) Novel insecticidal toxins from the venom of the spider Segestria florentina. Toxicon 40, 125–130.
Pluzhnikov, K., Vassilevski, A., Korolkova, Y. et al. (2007) Omega-Lsp-IA, a novel modulator of P-type Ca2+ channels. Toxicon 50, 993–1004.
Escoubas, P., Quinton, L., and Nicholson, G.M. (2008) Venomics: unravelling the complexity of animal venoms with mass spectrometry. J. Mass Spectrom. 43, 279–295.
Liang, S. (2008) Proteome and peptidome profiling of spider venoms. Expert Rev. Proteomics 5, 731–746.
Kozlov, S.A., Vassilevski, A.A., and Grishin, E.V. (2008) in “Peptidomics: Methods and Applications” (Soloviev, M., Shaw, C. and Andrén, P., Eds.), John Wiley & Sons, Hoboken, NJ, pp. 55–70.
Cutler, P. (2004) Protein purification protocols, Humana Press, Totowa, NJ.
Darbre, A. (1986) Practical protein chemistry: a handbook, Wiley, Chichester, NY.
Smith, B.J. (2003) Protein sequencing protocols, Humana Press, Totowa, NJ.
Otvos, L. and Cudic, M. (2007) Broth microdilution antibacterial assay of peptides. Methods Mol. Biol. 386, 309–320.
Wiegand, I., Hilpert, K., and Hancock, R.E. (2008) Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat. Protoc. 3, 163–175.
Acknowledgments
This work was funded by the Russian Foundation for Basic Research (Grant Nos. 08-04-00454 and 08-04-90444), the Federal Agency for Education of the Russian Federation (State Contact No. P 1388) and the Program of Cell and Molecular Biology of the Russian Academy of Sciences.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Vassilevski, A.A., Kozlov, S.A., Egorov, T.A., Grishin, E.V. (2010). Purification and Characterization of Biologically ctive Peptides from Spider Venoms. In: Soloviev, M. (eds) Peptidomics. Methods in Molecular Biology, vol 615. Humana Press. https://doi.org/10.1007/978-1-60761-535-4_7
Download citation
DOI: https://doi.org/10.1007/978-1-60761-535-4_7
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60761-534-7
Online ISBN: 978-1-60761-535-4
eBook Packages: Springer Protocols