[en] 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 (B. jararaca and C. rhodostoma, respectively) or three (E. ocellatus) active proteases were identified. Therefore, the assay allows the study of proteolytic activity towards the ECM in vitro for crude and fractionated venoms.
Disciplines :
Chemistry
Author, co-author :
Wachtel, Eric ; AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands
Bittenbinder, Matyas A; AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands ; Centre for Analytical Sciences Amsterdam (CASA), 1098 XH Amsterdam, The Netherlands ; Naturalis Biodiversity Center, Darwinweg 2, 2333 CR Leiden, The Netherlands
van de Velde, Bas; AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands ; Centre for Analytical Sciences Amsterdam (CASA), 1098 XH Amsterdam, The Netherlands
Slagboom, Julien; AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands ; Centre for Analytical Sciences Amsterdam (CASA), 1098 XH Amsterdam, The Netherlands
de Monts de Savasse, Axel ; Université de Liège - ULiège > Faculté de Médecine > Master sc. pharma., à fin. ; AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands
Alonso, Luis L ; AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands ; Centre for Analytical Sciences Amsterdam (CASA), 1098 XH Amsterdam, The Netherlands
Casewell, Nicholas R ; Centre for Snakebite Research & Interventions, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK ; Centre for Drugs and Diagnostics, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK
Vonk, Freek J; AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands ; Centre for Analytical Sciences Amsterdam (CASA), 1098 XH Amsterdam, The Netherlands ; Naturalis Biodiversity Center, Darwinweg 2, 2333 CR Leiden, The Netherlands
Kool, Jeroen ; AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands ; Centre for Analytical Sciences Amsterdam (CASA), 1098 XH Amsterdam, The Netherlands
Language :
English
Title :
Application of an Extracellular Matrix-Mimicking Fluorescent Polymer for the Detection of Proteolytic Venom Toxins.
COST - European Cooperation in Science and Technology Wellcome Trust
Funding number :
[221712/Z/20/Z] and [221710/Z/20/Z]
Funding text :
This research is in line with work from the COST Action European Venom Network CA19144, supported by COST (European Cooperation in Science and Technology) and was partly funded by the Wellcome Trust [221712/Z/20/Z] and [221710/Z/20/Z]. For the purpose of open access, the author has applied a CC BY public copyright license to any author accepted manuscript version arising from this submission.
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