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PreprintCited by 2Venomous Animal Envenomation and StudiesAmphibian and Reptile BiologyProtease and Inhibitor Mechanisms

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Purified Zymogens Reveal Mechanisms of Snake Venom Metalloproteinase Auto-Activation

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Hall, Sophie, Cardoso, Iara Aimê, Wilkinson, Mark C., Carretero, Maria Molina, Lingappa, Srikanth, Rand, Bronwyn, Shen, Dakang, Boldrini-França, Johara, Stenner, Richard, Menzies, Stefanie K., Balchin, Georgia, Hus, Konrad Kamil, Vincentelli, Renaud, Mumford, Andrew, Poole, Alastair W., Casewell, Nicholas R., Berger, Imre, Schaffitzel, Christiane·2025·eLife
DOI 10.1101/2025.06.27.661748

Preprint — published as 10.7554/elife.109112.3

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Abstract

Snake venoms contain diverse mixtures of toxins that evolved to incapacitate prey, but in humans they cause extensive pathology following snakebite envenomation. In viper venom, some of the most potent toxins are the haemorrhagic and coagulopathic snake venom metalloproteinases (SVMPs). Because venoms contain a SVMP cocktail, and due to their cytotoxicity, SVMP characterizations have been hampered by the lack of purified enzymes. By incorporating their prodomain, which blocks the active SVMP site, we overcame their cytotoxicity and enabled recombinant production of zymogens from all three structurally variable SVMP classes (PI, PII and PIII) using our baculovirus/insect cell expression system. Zymogens were auto-activated by incubation with Zn 2+ ions, resulting in prodomain cleavage, PII disintegrin cleavage and PIII prodomain proteolysis. Auto-activated SVMPs were characterized using protein substrate degradation, platelet aggregation and blood coagulation assays, benchmarked to native venom-purified SVMP. Our recombinant zymogen production protocol is generically applicable for the expression of SVMPs, unlocking biomedical use in haematology, and discovery of novel snakebite therapeutics.

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Metadata

Authors
Hall, S., Cardoso, I. A., Wilkinson, M. C., Carretero, M. M., Lingappa, S., Rand, B., Shen, D., Boldrini-França, J., Stenner, R., Menzies, S. K., Balchin, G., Hus, K. K., Vincentelli, R., Mumford, A., Poole, A. W., Casewell, N. R., Berger, I., & Schaffitzel, C.
Affiliations
  • University of Bristol (GB)
  • Liverpool School of Tropical Medicine (GB)
  • Rzeszów University of Technology (PL)
  • Centre National de la Recherche Scientifique (FR)
Corresponding author
Schaffitzel, Christiane
ORCID
Venue
eLife
Publisher
openRxiv
Published
2025-06-30
Volume
15
ISSN
2050-084X (ISSN-L)
DOI
10.1101/2025.06.27.661748
OpenAlex
W7164214462
Cited by
2
References
2
Citations by year
2026: 2
FWCI
8.09
Type
journal article
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
Funders
  • European Council (899670, 101149867)
  • Biotechnology and Biological Sciences Research Council (BB/Y007581/1)
  • Engineering and Physical Sciences Research Council (EP/Z002613/1)
Topics
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Toxicology › Venomous Animal Envenomation and Studies
  • Life Sciences › Agricultural and Biological Sciences › Ecology, Evolution, Behavior and Systematics › Amphibian and Reptile Biology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Protease and Inhibitor Mechanisms
Keywords
snake venom metalloproteinases, recombinant enzyme expression, baculovirus-insect cell expression, viper venom, disintegrins, platelet aggregation, coagulation, snakebite

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