Abstract
The worrisome emergence of pathogens resistant to conventional drugs has stimulated the search for new classes of antimicrobial and antiparasitic agents from natural sources. Antimicrobial peptides (AMPs), acting through mechanisms that do not rely on the interaction with a specific receptor, provide new possibilities for the development of drugs against resistant organisms. This study sought to purify and proteomically characterize the antimicrobial and antiparasitic peptidomes of B. atrox and B. jararacussu snake venoms against Gram-positive (Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus—MRSA), Gram-negative (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae) bacteria, and the protozoan parasites Leishmania amazonensis and Plasmodium falciparum (clone W2, resistant to chloroquine). To this end, B. atrox and B. jararacussu venom peptides were purified by combination of 3 kDa cut-off Amicon® ultracentrifugal filters and reverse-phase high-performance liquid chromatography, and then identified by electrospray-ionization Ion-Trap/Time-of-Flight mass spectrometry. Fourteen distinct peptides, with masses ranging from 443.17 to 1383.73 Da and primary structure between 3 and 13 amino acid residues, were sequenced. Among them, 13 contained unique sequences, including 4 novel bradykinin-potentiating-like peptides (BPPs), and a snake venom metalloproteinase tripeptide inhibitor (SVMPi). Although commonly found in Viperidae venoms, except for Bax-12, the BPPs and SVMPi here reported had not been described in B. atrox and B. jararacussu venoms. Among the novel peptides, some exhibited bactericidal activity towards P. aeruginosa and S. aureus, had low hemolytic effect, and were devoid of antiparasitic activity. The identified novel antimicrobial peptides may be relevant in the development of new drugs for the management of multidrug-resistant Gram-negative and Gram-positive bacteria.
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Acknowledgements
The authors wish to gratefully acknowledge Dr. Juan J. Calvete for valuable comments and discussion. Authors wish also to thank Dr. Kayena Delaix Zaqueo for B. atrox venom extraction and Uecson Suendel and Paulo R. M. Sampaio for the B. jararacussu photograph. Special thanks to the group of Anemones (Claudia Siqueira, Jeane Moraes, Tainara Rodrigues e Rafaela Diniz), for all the help in this work. Thanks to colleagues Hugo Vigerelli, Douglas Mariano, Tiago Bispo, Rosimar Esquerdo, Gabriela Romina Barredo and Silvana Giudicessi for helping in various aspects of this work. Authors also express their gratitude to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq/MCTIC, Grant # 406385/2018 [DCP]), Instituto Nacional de Epidemiologia na Amazônia Ocidental (INCT), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES/MEC), Fundação Rondônia de Amparo ao Desenvolvimento das Ações Científicas e Tecnológicas de Pesquisa do Estado de Rondônia (FAPERO) and the Universidad de Buenos Aires (UBA) y Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) for financial support and Financiadora de Estudos e Projetos (FINEP) Grants # 01.12.0450.0 and 01.09.0278.04. DCP is a CNPq fellow (301974/2019-5). The authors thank the Program for Technological Development in Tools for Health-PDTIS-FIOCRUZ for allowing the use of its facilities. SAC is researcher of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).
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Conceptualization: CASC, LAC, AMS, and RGS. Data curation and Formal analysis: CASC, RDS, LAC, and AMS. Funding acquisition: LAC, AMS, and RGS. Investigation and Methodology: CASC, LAC, AMS, RDS, DCP, APAS, CBGT, NBM, SAC, and SLS. Writing and editing: CASC, LAC, AMS, RDS, SAC, SLS, CBGT, and DCP.
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The blood samples used for the tests received a favorable opinion from the Research Ethics Committee (CEP) under the number of Presentation Certificate for Ethical Appreciation (CAAE) 44899715.2.0000.0011.
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da Silva Caldeira, C.A., Diniz-Sousa, R., Pimenta, D.C. et al. Antimicrobial peptidomes of Bothrops atrox and Bothrops jararacussu snake venoms. Amino Acids 53, 1635–1648 (2021). https://doi.org/10.1007/s00726-021-03055-y
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DOI: https://doi.org/10.1007/s00726-021-03055-y