Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins
The cytotoxicity caused by snake venoms is a serious medical problem that greatly contributes to the morbidity observed in snakebite patients. The cytotoxic components found in snake venoms belong to a variety of toxin classes and may cause cytotoxic effects by targeting a range of molecular structu...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/15/4/294 |
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| author | Eric Wachtel Matyas A. Bittenbinder Bas van de Velde Julien Slagboom Axel de Monts de Savasse Luis L. Alonso Nicholas R. Casewell Freek J. Vonk Jeroen Kool |
| author_facet | Eric Wachtel Matyas A. Bittenbinder Bas van de Velde Julien Slagboom Axel de Monts de Savasse Luis L. Alonso Nicholas R. Casewell Freek J. Vonk Jeroen Kool |
| author_sort | Eric Wachtel |
| collection | DOAJ |
| description | The cytotoxicity caused by snake venoms is a serious medical problem that greatly contributes to the morbidity observed in snakebite patients. The cytotoxic components found in snake venoms belong to a variety of toxin classes and may cause cytotoxic effects by targeting a range of molecular structures, including cellular membranes, the extracellular matrix (ECM) and the cytoskeleton. Here, we present a high-throughput assay (384-well plate) that monitors ECM degradation by snake venom toxins via the application of fluorescent versions of model ECM substrates, specifically gelatin and collagen type I. Both crude venoms and fractionated toxins of a selection of medically relevant viperid and elapid species, separated via size-exclusion chromatography, were studied using the self-quenching, fluorescently labelled ECM–polymer substrates. The viperid venoms showed significantly higher proteolytic degradation when compared to elapid venoms, although the venoms with higher snake venom metalloproteinase content did not necessarily exhibit stronger substrate degradation than those with a lower one. Gelatin was generally more readily cleaved than collagen type I. In the viperid venoms, which were subjected to fractionation by SEC, two (<i>B. jararaca</i> and <i>C. rhodostoma</i>, respectively) or three (<i>E. ocellatus</i>) active proteases were identified. Therefore, the assay allows the study of proteolytic activity towards the ECM in vitro for crude and fractionated venoms. |
| first_indexed | 2024-03-11T04:28:32Z |
| format | Article |
| id | doaj.art-df8a65ad150346649043fa585e91e25c |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-11T04:28:32Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-df8a65ad150346649043fa585e91e25c2023-11-17T21:39:09ZengMDPI AGToxins2072-66512023-04-0115429410.3390/toxins15040294Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom ToxinsEric Wachtel0Matyas A. Bittenbinder1Bas van de Velde2Julien Slagboom3Axel de Monts de Savasse4Luis L. Alonso5Nicholas R. Casewell6Freek J. Vonk7Jeroen Kool8AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsAIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsAIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsAIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsAIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsAIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsCentre for Snakebite Research & Interventions, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UKAIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsAIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsThe cytotoxicity caused by snake venoms is a serious medical problem that greatly contributes to the morbidity observed in snakebite patients. The cytotoxic components found in snake venoms belong to a variety of toxin classes and may cause cytotoxic effects by targeting a range of molecular structures, including cellular membranes, the extracellular matrix (ECM) and the cytoskeleton. Here, we present a high-throughput assay (384-well plate) that monitors ECM degradation by snake venom toxins via the application of fluorescent versions of model ECM substrates, specifically gelatin and collagen type I. Both crude venoms and fractionated toxins of a selection of medically relevant viperid and elapid species, separated via size-exclusion chromatography, were studied using the self-quenching, fluorescently labelled ECM–polymer substrates. The viperid venoms showed significantly higher proteolytic degradation when compared to elapid venoms, although the venoms with higher snake venom metalloproteinase content did not necessarily exhibit stronger substrate degradation than those with a lower one. Gelatin was generally more readily cleaved than collagen type I. In the viperid venoms, which were subjected to fractionation by SEC, two (<i>B. jararaca</i> and <i>C. rhodostoma</i>, respectively) or three (<i>E. ocellatus</i>) active proteases were identified. Therefore, the assay allows the study of proteolytic activity towards the ECM in vitro for crude and fractionated venoms.https://www.mdpi.com/2072-6651/15/4/294snakebitetoxinsSVMPextracellular matrixcytotoxicitytissue damage |
| spellingShingle | Eric Wachtel Matyas A. Bittenbinder Bas van de Velde Julien Slagboom Axel de Monts de Savasse Luis L. Alonso Nicholas R. Casewell Freek J. Vonk Jeroen Kool Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins Toxins snakebite toxins SVMP extracellular matrix cytotoxicity tissue damage |
| title | Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins |
| title_full | Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins |
| title_fullStr | Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins |
| title_full_unstemmed | Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins |
| title_short | Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins |
| title_sort | application of an extracellular matrix mimicking fluorescent polymer for the detection of proteolytic venom toxins |
| topic | snakebite toxins SVMP extracellular matrix cytotoxicity tissue damage |
| url | https://www.mdpi.com/2072-6651/15/4/294 |
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