Deciphering the venom components of Sea snakes: Integrating dual proteomic techniques to uncover the bioactive potential of Hydrophis curtus.

Proteomics constitutes a crucial technology in marine evolutionary biology and the discovery of novel natural drugs. Spine-bellied Sea Snake (Hydrophis curtus), noted for its extensive distribution and highly potent venom, offers exceptional opportunities for scientific exploration and therapeutic innovation. While preliminary insights into the venom's active components have been acquired through conventional chromatography techniques, a comprehensive understanding of its protein architecture remains unclear. To advance this understanding, we employed tandem mass tagging (TMT) and data-independent acquisition (DIA) proteomics methodologies to perform an extensive and comparative analysis of H. curtus venom. The integration of these approaches facilitates a detailed illustration of the venom's composition, underscoring the specific advantages of each technique for diverse research objectives. Our study identifies several active substances within the Venom Factor (VF) family, suggesting their potential critical role in modulating the biological activity of venom proteins. A thorough investigation of the phospholipase A (PLA), three-finger toxins (3FTx), and cysteine-rich secretory proteins (CRISP) families present in the venom was undertaken. This study utilized structural simulations with AlphaFold2, comparative analyses using PyMOL, and phylogenetic reconstructions through MEGA-CC. These integrative methodologies elucidated the intricate molecular functions and underscored the potential for discovering novel bioactive compounds within the venom. Ultimately, this research provides an enhanced understanding of H. curtus venom, laying a robust work for future studies on toxin evolution and the development of marine-derived drugs.