Australian scorpion Hormurus waigiensis venom fractions show broad bioactivity through modulation of bio-impedance and cytosolic calcium
Housley, David M., Pinyon, Jeremy L., Jonquieres, Georg von, Perera, Chamini J., Smout, Michael, Liddell, Michael J., Jennings, Ernest A., Wilson, David, and Housley, Gary D. (2020) Australian scorpion Hormurus waigiensis venom fractions show broad bioactivity through modulation of bio-impedance and cytosolic calcium. Biomolecules, 10 (4). 617.
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Abstract
Scorpion venoms are a rich source of bioactive molecules, but characterisation of toxin peptides affecting cytosolic Ca2+, central to cell signalling and cell death, is limited. We undertook a functional screening of the venom of the Australian scorpion Hormurus waigiensis to determine the breadth of Ca2+ mobilisation. A human embryonic kidney (HEK293) cell line stably expressing the genetically encoded Ca2+ reporter GCaMP5G and the rabbit type 1 ryanodine receptor (RyR1) was developed as a biosensor. Size-exclusion Fast Protein Liquid Chromatography separated the venom into 53 fractions, constituting 12 chromatographic peaks. Liquid chromatography mass spectroscopy identified 182 distinct molecules with 3 to 63 components per peak. The molecular weights varied from 258 Da—13.6 kDa, with 53% under 1 kDa. The majority of the venom chromatographic peaks (tested as six venom pools) were found to reversibly modulate cell monolayer bioimpedance, detected using the xCELLigence platform (ACEA Biosciences). Confocal Ca2+ imaging showed 9/14 peak samples, with molecules spanning the molecular size range, increased cytosolic Ca2+ mobilization. H. waigiensis venom Ca2+ activity was correlated with changes in bio-impedance, reflecting multi-modal toxin actions on cell physiology across the venom proteome.
| Item ID: | 63476 |
|---|---|
| Item Type: | Article (Research - C1) |
| ISSN: | 2218-273X |
| Keywords: | membrane biophysics; scorpion toxins; xCELLigence Real Time Cell Analysis; GCaMP5Gcalcium reporter; ryanodine receptors; calcium store; HEK293 cells; scorpion envenomation; recombinant rabbit RyR1; Ca2+ biosensor |
| Copyright Information: | © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license. |
| Funders: | James Cook University, University of New South Wales |
| Date Deposited: | 22 Jul 2020 06:21 |
| FoR Codes: | 31 BIOLOGICAL SCIENCES > 3101 Biochemistry and cell biology > 310112 Structural biology (incl. macromolecular modelling) @ 50% 32 BIOMEDICAL AND CLINICAL SCIENCES > 3214 Pharmacology and pharmaceutical sciences > 321499 Pharmacology and pharmaceutical sciences not elsewhere classified @ 50% |
| SEO Codes: | 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100% |
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