RT info:eu-repo/semantics/article
T1 Cystatin from Austrelaps superbus snake venom as a model for identifying potential inhibitors of Trypanosoma cruzi cruzain
A1 Alfonso Ruiz Díaz, Jorge Javier
A1 Gómez Garay, Ana Fidelina
A1 Makoto Kayano, Anderson
A1 Holanda, Rudson
A1 Ferreira Francisco, Aleff
A1 Collins Kuehn, Christian
A1 Martins Soares, Andreimar
A1 Vega Gómez, María Celeste
A1 de Azevedo Calderon, Leonardo
AB Background: Chagas disease (CD), caused by Trypanosoma cruzi, affects approximately seven million individuals worldwide, with the highest number of cases in Latin America. CD has two phases, of which the chronic phase is characterized by reduced efficacy in drug therapies. This and other factors make developing new strategies that aim to identify molecules capable of becoming alternatives to or complement current chemotherapy vitally important.Methods: Cruzain and AsCystatin were obtained recombinantly through expression in E. coli. Bioinformatic assays were conducted with both molecules, followed by in vitro enzyme inhibition assays. Subsequently, in silico studies allowed for the design of peptides, which were then assessed for molecular interactions with cruzain. The designed peptides were synthesized, and their inhibitory potential on cruzain and their trypanocidal and cytotoxic effects in vitro were finally assessed.Results: AsCystatin, a potential inhibitor of cysteine proteases, was identified from previously published scientific literature. In silico assays suggested that AsCystatin interacts with key regions of cruzain, and was subsequently produced through heterologous expression, obtaining a protein with a high degree of purity. Next, the inhibition of AsCystatin on the activity of cruzain was assessed, observing that approximately 20 μM of cystatin could inhibit 50% of the catalytic activity of the recombinant enzyme. Based on the in-silico analysis performed previously, original, and modified peptides were designed and tested, which allowed for identifying four peptides with inhibitory capacity on the enzymatic activity of cruzain. Finally, three of these peptides showed trypanocidal activity on epimastigote forms of T. cruzi in in vitro models.Conclusion: It was possible to identify AsCystatin and four peptides derived from this protein with inhibitory activity on cruzain, highlighting the trypanocidal effect of these peptides observed in in vitro assays.
PB Universidade Estadual Paulista Júlio de Mesquita Filho. Centro de Estudos de Venenos e Animais Peçonhentos
YR 2025
FD 2025-02-14
LK http://hdl.handle.net/20.500.14066/4647
UL http://hdl.handle.net/20.500.14066/4647
LA eng
NO Alfonso Ruiz Díaz, J. J., Gómez Garay, A. F., Makoto Kayano, A., Holanda, R., Ferreira Francisco, A., Collins Kuehn, C., Martins Soares, A., Vega, C., & de Azevedo Calderon, L. (2025). Cystatin from Austrelaps superbus snake venom as a model for identifying potential inhibitors of Trypanosoma cruzi cruzain. Journal of Venomous Animals and Toxins including Tropical Diseases, 31, Article e20240055. https://doi.org/10.1590/1678-9199-JVATITD-2024-0055
NO Correspondence: calderon@unir.br.
NO Consejo Nacional de Ciencia y Tecnología
DS MINDS@UW
RD 03-sep-2026