Structural, biochemical and immunochemical characterization of an acidic phospholipase A2 from Lachesis acrochorda (Viperidae: Crotalinae) venom

• Published in: Toxicon (Oxford) Vol. 237; p. 107528
• Main Authors: Franco-Vásquez, Adrián Marcelo, Lazcano-Pérez, Fernando, Mejía-Sánchez, Miguel Angel, Corzo, Gerardo, Zamudio, Fernando, Carbajal-Saucedo, Alejandro, Román-González, Sergio Agustín, Gómez-Manzo, Saúl, Arreguín-Espinosa, Roberto
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.01.2024
• Subjects: Antibody; Bushmasters; Lachesis acrochorda; Mass spectrometry; Phospholipase A2; Venom; Phospholipase A2; Antibody; Mass spectrometry; Lachesis acrochorda; Venom; Bushmasters; Phospholipase A
• ISSN: 0041-0101; 1879-3150
• DOI: 10.1016/j.toxicon.2023.107528

Viperids of the genus Lachesis, also known as bushmasters, are capable of injecting great amounts of venom that cause severe envenomation incidents. Since phospholipases type A2 are mainly involved in edema and myonecrosis within the snakebite sites, in this work, the isolation, amino acid sequence and biochemical characterization of the first phospholipase type A2 from the venom of Lachesis acrochorda, named Lacro_PLA2, is described. Lacro_PLA2 is an acidic aspartic 49 calcium-dependent phospholipase A2 with 93% similarity to the L. stenophrys phospholipase. Lacro_PLA2 has a molecular mass of 13,969.7 Da and an experimental isoelectric point around 5.3. A combination of N-terminal Edman degradation and MS/MS spectrometry analyses revealed that Lacro_PLA2 contains 122 residues including 14 cysteines that form 7 disulfide bridges. A predicted 3D model shows a high resemblance to other viperid phospholipases. Nevertheless, immunochemical and phospholipase neutralization tests revealed a notorious level of immunorecognition of the isolated protein by two polyclonal antibodies from viperids from different genus, which suggest that Lacro_PLA2 resembles more to bothropic phospholipases. Lacro_PLA2 also showed significantly high edema activity when was injected into mice; so, it could be an alternative antigen in the development of antibodies against toxins of this group of viperids, seeking to improve commercial polyclonal antivenoms. [Display omitted] •An acidic phospholipase A2 (PLA2) was isolated from the venom of the bushmaster snake Lachesis acrochorda•The amino acid sequence and predicted 3D structure were determined and showed high homology with other PLA2S from viperids•Mass spectrometry analysis determined that the protein is an aspartic 49 PLA2 with catalytic activity and calcium-dependency•The immunochemical tests revealed a high level of recognition by two polyclonal antibodies from different viperid PLA2s•The phospholipase is not lethal to mice at the maximum doses tested (30 μg) but exerts considerable edema activity