The double-banded false coralsnake Erythrolamprus bizona (Dipsadidae, Xenodontinae, Xenodontini) has a metalloproteinase-rich venom with proteolytic activity towards azocasein and α-fibrinogen

Article

作者: Bayona-Serrano, Juan D ; Muñoz-Gómez, Luis M ; Bernal-Bautista, Manuel H ; Torres-Bonilla, Kristian A ; Sáenz-Suarez, Paula A ; Hyslop, Stephen

The venom of the double-banded false coralsnake, Erythrolamprus bizona, is proteolytic and attenuates neuromuscular contractile activity in vitro. Here, we examined the Duvernoy's venom gland histology and general composition of E. bizona venom using a combination of chromatographic, electrophoretic, enzymatic and proteomic analyses. Histologically, the venom gland consisted of serous epithelium-lined secretory tubules and a supralabial gland that stained positively for mucopolysaccharide. SDS-PAGE showed that the venom had a simple composition, with proteins in the range of 15-60 kDa. This simple composition was confirmed by RP-HPLC that revealed 15 main protein peaks. The venom (1-10 μg) was highly proteolytic towards azocasein, but was devoid of esterase, phospholipase (PLA₂), and L-amino acid oxidase activities. The venom also degraded casein and gelatin in zymographic assays, with activity towards gelatin being particularly potent and detected over the range of 18.7 ng-30 μg of venom; gelatinolytic activity was also detected in four of the RP-HPLC peaks. The venom (10 μg) selectively degraded the α-chain of fibrinogen. All proteolytic activity was inhibited by EDTA (metalloproteinase inhibitor) but not by AEBSF (serine proteinase inhibitor). SDS-PAGE followed by in-gel digestion of the main electrophoretic bands coupled with LC-MS/MS analysis revealed the presence of five toxin families: C-type lectin-like proteins (CTL), cysteine-rich secretory proteins (CRiSP), phospholipase B (PLB), snake venom matrix metalloproteinases (svMMP), and snake venom metalloproteinases (SVMP). These findings extend our knowledge of the toxinology of E. bizona and suggest that the local manifestations (pain, edema, erythema, and ecchymosis) seen in human envenomation by this species are probably mediated by venom metalloproteinases.

2024-05-01·Archives of toxicology

Role of the complement system in kidney cell death induced by Loxosceles venom Sphingomyelinases D

Article

作者: Okamoto, Cinthya Kimori ; Tambourgi, Denise V ; van den Berg, Carmen W ; Pohl, Paula C

Envenomation by Loxosceles spiders can result in local and systemic pathologies. Systemic loxoscelism, which can lead to death, is characterized by intravascular hemolysis, platelet aggregation, and acute kidney injury. Sphingomyelinase D (SMase D) in Loxosceles spider venom is responsible for both local and systemic pathologies, and has been shown to induce metalloprotease activity. As the complement system is involved in many renal pathologies and is involved in hemolysis in systemic loxoscelism, the aim of this study was to investigate its role and the role of complement regulators and metalloproteases in an in vitro model of Loxosceles venom induced renal pathology. We investigated the effects of the venom/SMase D and the complement system on the HK-2 kidney cell line. Using cell viability assays, western blotting, and flow cytometry, we show that human serum, as a source of complement, enhanced the venom/SMase D induced cell death and the deposition of complement components and properdin. Inhibitors for ADAM-10 and ADAM-17 prevented the venom induced release of the of the complement regulator MCP/CD46 and reduced the venom/SMase D induced cell death. Our results show that the complement system can contribute to Loxosceles venom induced renal pathology. We therefore suggest that patients experiencing systemic loxoscelism may benefit from treatment with metalloproteinase inhibitors and complement inhibitors, but this proposition should be further analyzed in future pre-clinical and clinical assays.

2024-04-01·CURRENT OPINION IN BIOTECHNOLOGY

Engineering metalloproteinase inhibitors: tissue inhibitors of metalloproteinases or antibodies, that is the question

Review

作者: Hilpert, Gregory A ; Raeeszadeh-Sarmazdeh, Maryam ; Mosca, Ethan R ; Kalantar, Masoud

Targeting metalloproteinases (MPs) has been the center of attention for developing therapeutics due to their contribution to a wide range of diseases, including cancer, cardiovascular, neurodegenerative disease, and preterm labor. Protein-based MP inhibitors offer higher stability and selectivity, which is critical for developing efficient therapeutics with low off-target effects. Tissue inhibitors of metalloproteinases (TIMPs), natural inhibitors of MPs, and antibodies provide excellent protein scaffolds for engineering selective or multispecific MP inhibitors. Advances in protein engineering and design techniques, such as rational design and directed evolution using yeast display to develop potent MP inhibitors, are discussed, including but not limited to loop grafting, swapping, and counterselective selection.

100 项与 01-07A 相关的药物交易

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