Evidence of venom variability associated with venom extraction interval in the monocled cobra (Naja kaouthia)

  Evidence of venom variability associated with venom extraction interval in the monocled cobra (Naja kaouthia) Abstract Snake venom variability is a critical factor in antivenom production. Although several influencing factors have been studied, the impact of extraction interval on venom properties remains poorly understood. This study evaluated the effects of various extraction intervals on venom yield, the percentage of dry weight (%Dw), and composition in the monocled cobra ( Naja kaouthia ). Twenty-five captive snakes were randomly allocated to five groups, each undergoing a control extraction followed by a repeated extraction at 1, 4, 7, 10, or 14 days. Venom profiles were characterized using reversed-phase high-performance liquid chromatography (RP-HPLC) and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). Results indicated that while fresh venom yield remained stable across all intervals, %Dw significantly decreased in extractions performed between 1 and 10 days, ...

Pioneering Comparative Proteomic and Enzymatic Profiling of Amazonian Scorpion Venoms Enables the Isolation of Their First α-Ktx, Metalloprotease, and Phospholipase A2

 


Pioneering Comparative Proteomic and Enzymatic Profiling of Amazonian Scorpion Venoms Enables the Isolation of Their First α-Ktx, Metalloprotease, and Phospholipase A2

Abstract

Scorpionism is a growing public health concern in Brazil, with the Amazon region presenting the highest mortality rates but remaining understudied, especially regarding local scorpion venoms composition. This study presents the first comprehensive biochemical characterization of venoms from three Amazonian species—Tityus metuendus (TmetuV), Tityus silvestris (TsilvV), and Brotheas amazonicus (BamazV)—using an integrated approach combining Multi-Enzymatic Limited Digestion (MELD)-based bottom-up proteomics, high-resolution LC-MS/MS, chromatography, zymography, and enzymatic assays. Tityus serrulatus venom was included as a reference. Significant biochemical differences were observed: TsilvV was rich in 20–30 kDa proteins and showed strong metalloprotease activity; BamazV exhibited high molecular weight proteins and potent phospholipase A2 (PLA2) activity but lacked proteolytic and fibrinogenolytic activities; TmetuV showed the highest hyaluronidase activity and abundance of α-KTx neurotoxins. Zymography revealed a conserved ~45 kDa hyaluronidase in all species. Three novel components were partially characterized: BamazPLA2 (Group III PLA2), Tmetu1 (37-residue α-KTx), and TsilvMP_A (a metalloprotease homologous to antarease). This is the first application of MELD-based proteomics to Amazonian scorpion venoms, revealing molecular diversity and functional divergence within Tityus and Brotheas, emphasizing the need for region-specific antivenoms. These findings provide a foundation for future pharmacological studies and the discovery of bioactive peptides with therapeutic potential.

Bordon KCF, Santos GC, Martins JG, Wiezel GA, Amorim FG, Crasset T, Redureau D, Quinton L, Procópio REL, Arantes EC. Pioneering Comparative Proteomic and Enzymatic Profiling of Amazonian Scorpion Venoms Enables the Isolation of Their First α-Ktx, Metalloprotease, and Phospholipase A2. Toxins. 2025; 17(8):411. https://doi.org/10.3390/toxins17080411