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Comprehensive Proteome Reveals the Key Lethal Toxins in the Venom of Jellyfish Nemopilema nomurai.

Jellyfish stings are a major threat to human beings in coastal areas of the world. Each year, hundreds of thousands of victims are stung by venomous jellyfish. Nemopilema nomurai is a dangerous species with a large number of victims including many deaths. N. nomurai venom is a complex cocktail that is rich in proteins and peptides, and it is secreted by nematocysts for prey or defense. Previous studies have identified hundreds of toxins in the venom of N. nomurai; however, it is unclear which toxin(s) is responsible for lethality. Herein, we isolated the lethal fraction (NnLF) from N. nomurai venom with multiple chromatography. NnLF showed strong lethality to mice, and the toxicology results were consistent with the clinical symptoms of dead patients after N. nomurai sting, which indicated that NnLF contained the key lethal toxins in the venom. Subsequently, proteomic analysis was performed to identify the toxins in NnLF, and a total of 13 toxin homologs were identified, including phospholipase, potassium channel inhibitor, hemolysin, thrombin, etc. Moreover, in vitro toxicity assays further verified the phospholipase A2 and hemolytic activity of NnLF. These results revealed that cell membrane-targeted toxins, including channel-forming toxins, potassium channel inhibitors, and especially phospholipases, played very important roles in the lethality of N. nomurai sting. Moreover, blood toxins such as thrombin-like toxin and hemolysins might be synergistically involved in lethality. These findings advance the understanding of lethality caused by N. nomurai sting and will be significant for the development of drugs to treat this jellyfish sting in the future.