Abstract
For medical use of proteins and peptide-based drugs, it is desirable to have small biologically active sequences because they improve stability, reduce side effects, and production costs. Several plant defensins have their biological activities imparted by a sequence named γ-core. Vu-Def, a Vigna unguiculata defensin, has activity against Leishmania amazonensis, which is one etiological agent of leishmaniasis and for which new drugs are needed. Our intention was to understand if the region comprising the Vu-Def γ-core is responsible for the biological activity against L. amazonensis and to unveil its mechanism of action. Different microbiological assays with L. amazonensis in the presence of the synthetic peptide A36,42,44γ32-46Vu-Def were done, as well as ultrastructural and fluorescent analyses. A36,42,44γ32-46Vu-Def showed biological activity similar to Vu-Def. A36,42,44γ32-46Vu-Def (74 µM) caused 97% inhibition of L. amazonensis culture and parasites were unable to regrow in fresh medium. The cells of the treated parasites showed morphological alterations by ultrastructural analysis and fluorescent labelings that corroborate with the data of the organelles alterations. The general significance of our work is based on the description of a small synthetic peptide, A36,42,44γ32-46Vu-Def, which has activity on L. amazonensis and that the interaction between A36,42,44γ32-46Vu-Def-L. amazonensis results in parasite inhibition by the activation of an apoptotic-like cell death pathway.
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Acknowledgements
We acknowledge Luiz Carlos Dutra de Souza and Valéria Miguelote Kokis for their technical support. This work has received financial support of the Brazilian agencies Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq), Fundação de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ, process no. E-26/102.981/2012-Bolsa; process no. E-26/202.760/2018-Bolsa), and the support of Universidade Estadual do Norte Fluminense. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-Brasil (CAPES)-Finance Code 001.
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GSS and LPC performed the experiments; FCVS prepared the modeling; AOC and OLTM participated in the peptide design; EJTM, VMG and AOC participated in the protozoan experimental design and data analysis; GSS and AOC contributed to the writing of the manuscript.
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Souza, G.S., de Carvalho, L.P., de Melo, E.J.T. et al. A synthetic peptide derived of the β2–β3 loop of the plant defensin from Vigna unguiculata seeds induces Leishmania amazonensis apoptosis-like cell death. Amino Acids 51, 1633–1648 (2019). https://doi.org/10.1007/s00726-019-02800-8
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DOI: https://doi.org/10.1007/s00726-019-02800-8