Proteomic profiling and comparative hemotoxicity of Gloydius brevicaudus and Deinagkistrodon acutus venoms: insights into functional convergence

Abstract

Background: Snakebite envenomation caused by G. brevicaudus and D. acutus frequently presents with overlapping hemorrhagic and coagulopathic manifestations in clinical settings. However, whether these similarities reflect convergent venom phenotypes remains unclear. Methods: We performed an integrated comparative analysis combining nano-LC-MS/MS proteomics with multi-target functional assays to systematically evaluate venom composition and biological activities. Proteomic profiling was used to characterize toxin family abundance, while enzymatic, cytotoxic, hemolytic, and cell signaling assays were conducted to assess functional effects. Results: Proteomic analysis revealed that both venoms are predominantly composed of hemotoxic toxin families, including snake venom metalloproteinases (SVMP), snake venom serine proteases (SVSP), C-type lectin-like proteins (CTLP), and phospholipase A₂ (PLA₂), with broadly comparable relative abundances. Functional assays demonstrated similar concentration-dependent patterns in proteolytic, PLA₂, thrombin-like, and fibrinolytic activities, with no statistically significant interspecific differences. Both venoms also induced comparable cytotoxic effects across mammalian cell lines, while exhibiting limited hemolytic activity and minimal modulation of Ca²⁺ signaling and nitric oxide production. Conclusions: These findings demonstrate that G. brevicaudus and D. acutus venoms share a convergent hemotoxic functional architecture characterized by consistent enzymatic activities and similar cytotoxic profiles. This functional convergence provides a mechanistic basis for clinically overlapping hemorrhagic and coagulopathic manifestations observed in envenomation cases. The results further emphasize the importance of function-oriented venom profiling and suggest potential implications for antivenom cross-reactivity and therapeutic development.