Top-down proteomics of venoms from five Micrurus species from Costa Rica: comparative composition of phospholipase A2-rich vs three-finger toxin-rich phenotypes

Coralsnakes of the genus Micrurus include more than 80 species distributed in the American continent. They produce potent neurotoxic venoms acting at the neuromuscular junction and potentially leading to respiratory paralysis and death. The vast majority of proteins in coralsnake venoms belong to the three-finger toxin (3FTx) and the group I phospholipase A₂ (PLA₂) families. Previous studies using ‘bottom-up’ proteomic strategies have revealed a compositional dichotomy of toxin expression by which different Micrurus species display a predominance of either 3FTx or PLA₂ proteins in their venoms, possibly linked to the phylogeographic structure of the genus radiation. ‘Top-down’ proteomics (TDP) allows the direct analysis of intact proteins in a high resolution mass spectrometer, circumventing the limitations of the ‘peptide-to-protein inference problem’ inherent to the bottom-up approach. Here, we analyzed the venoms of five out of the six Micrurus species that inhabit Costa Rica, by using a TDP approach. Results unveil venom proteoforms that are shared between these species, and provide additional insights into the variable compositional complexity of these venoms and relationships to their 3FTx/PLA₂ dichotomy.