Australian Scorpion <i>Hormurus waigiensis</i> Venom Fractions Show Broad Bioactivity through Modulation of Bio-Impedance and Cytosolic Calcium
Scorpion venoms are a rich source of bioactive molecules, but characterisation of toxin peptides affecting cytosolic Ca<sup>2+</sup>, central to cell signalling and cell death, is limited. We undertook a functional screening of the venom of the Australian scorpion <i>Hormurus waigi...
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| Format: | Article |
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MDPI AG
2020-04-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/4/617 |
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| author | David M. Housley Jeremy L. Pinyon Georg von Jonquieres Chamini J. Perera Michael Smout Michael J. Liddell Ernest A. Jennings David Wilson Gary D. Housley |
| author_facet | David M. Housley Jeremy L. Pinyon Georg von Jonquieres Chamini J. Perera Michael Smout Michael J. Liddell Ernest A. Jennings David Wilson Gary D. Housley |
| author_sort | David M. Housley |
| collection | DOAJ |
| description | Scorpion venoms are a rich source of bioactive molecules, but characterisation of toxin peptides affecting cytosolic Ca<sup>2+</sup>, central to cell signalling and cell death, is limited. We undertook a functional screening of the venom of the Australian scorpion <i>Hormurus waigiensis</i> to determine the breadth of Ca<sup>2+</sup> mobilisation. A human embryonic kidney (HEK293) cell line stably expressing the genetically encoded Ca<sup>2+</sup> 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 Ca<sup>2+</sup> imaging showed 9/14 peak samples, with molecules spanning the molecular size range, increased cytosolic Ca<sup>2+</sup> mobilization. <i>H. waigiensis</i> venom Ca<sup>2+</sup> activity was correlated with changes in bio-impedance, reflecting multi-modal toxin actions on cell physiology across the venom proteome. |
| first_indexed | 2024-03-10T20:26:20Z |
| format | Article |
| id | doaj.art-a13ed9de228c4168badb65fd4b07b9a5 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T20:26:20Z |
| publishDate | 2020-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-a13ed9de228c4168badb65fd4b07b9a52023-11-19T21:48:32ZengMDPI AGBiomolecules2218-273X2020-04-0110461710.3390/biom10040617Australian Scorpion <i>Hormurus waigiensis</i> Venom Fractions Show Broad Bioactivity through Modulation of Bio-Impedance and Cytosolic CalciumDavid M. Housley0Jeremy L. Pinyon1Georg von Jonquieres2Chamini J. Perera3Michael Smout4Michael J. Liddell5Ernest A. Jennings6David Wilson7Gary D. Housley8Translational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Sydney, Sydney, NSW 2052, AustraliaTranslational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Sydney, Sydney, NSW 2052, AustraliaTranslational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Sydney, Sydney, NSW 2052, AustraliaTranslational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Sydney, Sydney, NSW 2052, AustraliaAustralian Institute of Tropical Health and Medicine, James Cook University, Cairns, QLD 4878, AustraliaCentre for Tropical Environmental and Sustainability Science, College of Science & Engineering, Cairns Campus, James Cook University, Cairns, QLD 4878, AustraliaCollege of Medicine and Dentistry, Cairns Campus, James Cook University, Cairns, QLD 4878, AustraliaAustralian Institute of Tropical Health and Medicine, James Cook University, Cairns, QLD 4878, AustraliaTranslational Neuroscience Facility and Department of Physiology, School of Medical Sciences, UNSW Sydney, Sydney, NSW 2052, AustraliaScorpion venoms are a rich source of bioactive molecules, but characterisation of toxin peptides affecting cytosolic Ca<sup>2+</sup>, central to cell signalling and cell death, is limited. We undertook a functional screening of the venom of the Australian scorpion <i>Hormurus waigiensis</i> to determine the breadth of Ca<sup>2+</sup> mobilisation. A human embryonic kidney (HEK293) cell line stably expressing the genetically encoded Ca<sup>2+</sup> 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 Ca<sup>2+</sup> imaging showed 9/14 peak samples, with molecules spanning the molecular size range, increased cytosolic Ca<sup>2+</sup> mobilization. <i>H. waigiensis</i> venom Ca<sup>2+</sup> activity was correlated with changes in bio-impedance, reflecting multi-modal toxin actions on cell physiology across the venom proteome.https://www.mdpi.com/2218-273X/10/4/617membrane biophysicsscorpion toxinsxCELLigence Real Time Cell AnalysisGCaMP5G calcium reporterryanodine receptorscalcium store |
| spellingShingle | David M. Housley Jeremy L. Pinyon Georg von Jonquieres Chamini J. Perera Michael Smout Michael J. Liddell Ernest A. Jennings David Wilson Gary D. Housley Australian Scorpion <i>Hormurus waigiensis</i> Venom Fractions Show Broad Bioactivity through Modulation of Bio-Impedance and Cytosolic Calcium Biomolecules membrane biophysics scorpion toxins xCELLigence Real Time Cell Analysis GCaMP5G calcium reporter ryanodine receptors calcium store |
| title | Australian Scorpion <i>Hormurus waigiensis</i> Venom Fractions Show Broad Bioactivity through Modulation of Bio-Impedance and Cytosolic Calcium |
| title_full | Australian Scorpion <i>Hormurus waigiensis</i> Venom Fractions Show Broad Bioactivity through Modulation of Bio-Impedance and Cytosolic Calcium |
| title_fullStr | Australian Scorpion <i>Hormurus waigiensis</i> Venom Fractions Show Broad Bioactivity through Modulation of Bio-Impedance and Cytosolic Calcium |
| title_full_unstemmed | Australian Scorpion <i>Hormurus waigiensis</i> Venom Fractions Show Broad Bioactivity through Modulation of Bio-Impedance and Cytosolic Calcium |
| title_short | Australian Scorpion <i>Hormurus waigiensis</i> Venom Fractions Show Broad Bioactivity through Modulation of Bio-Impedance and Cytosolic Calcium |
| title_sort | australian scorpion i hormurus waigiensis i venom fractions show broad bioactivity through modulation of bio impedance and cytosolic calcium |
| topic | membrane biophysics scorpion toxins xCELLigence Real Time Cell Analysis GCaMP5G calcium reporter ryanodine receptors calcium store |
| url | https://www.mdpi.com/2218-273X/10/4/617 |
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