RESUMO
Antimicrobial peptides (AMP) are molecules with a broad spectrum of activities that have been identified in most living organisms. In addition, synthetic AMPs designed from natural polypeptides have been largely investigated. Here, we designed a novel AMP using the amino acid sequence of a plant trypsin inhibitor from Adenanthera pavonina seeds (ApTI) as a template. The 176 amino acid residues ApTI sequence was cleaved in silico using the Collection of Antimicrobial Peptides (CAMPR3), through the sliding-window method. Further improvements in AMP structure were carried out, resulting in adepamycin, an AMP designed from ApTI. Adepamycin showed antimicrobial activity from 0.9 to 3.6 µM against Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus strains. Moreover, this peptide also displayed activity against Candida albicans and Candida tropicalis. No toxic effects were observed on healthy human cells. Studies on the mechanism of action of adepamycin were carried out using an E. coli and C. tropicalis. Adepamycin triggers membrane disturbances, leading to intracellular nucleic acids release in E. coli. For C. tropicalis, an initial interference with the plasma membrane integrity is followed by the formation of intracellular reactive oxygen species (ROS), leading to apoptosis. Structurally, adepamycin was submitted to circular dichroism spectroscopy, molecular modeling and molecular dynamics simulations, revealing an environment-dependent α-helical structure in the presence of 2,2,2- trifluoroethanol (TFE) and in contact with mimetic vesicles/membranes. Therefore, adepamycin represents a novel lytic AMP with dual antibacterial and antifungal properties.
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Sequência de Aminoácidos , Antibacterianos/síntese química , Antibacterianos/toxicidade , Antifúngicos/síntese química , Antifúngicos/toxicidade , Peptídeos Catiônicos Antimicrobianos/síntese química , Peptídeos Catiônicos Antimicrobianos/toxicidade , Bactérias/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida tropicalis/efeitos dos fármacos , Membrana Celular/metabolismo , Permeabilidade da Membrana Celular/efeitos dos fármacos , Fabaceae/química , Hemólise/efeitos dos fármacos , Humanos , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Testes de Sensibilidade Microbiana , Simulação de Dinâmica Molecular , Fosfatidilgliceróis/químicaRESUMO
Hospital infection caused by Gram-negative bacteria is a serious and common problem, especially in developing countries. Aiming to reduce these infections, this report focuses on the identification and characterization of novel antimicrobial peptides from sesame (Sesamum indicum) kernel meals. Thus, sesame flour was extracted and precipitated with ammonium sulfate (100%). After dialysis, a rich fraction was applied to affinity red-Sepharose CL-6B chromatography, followed by reversed-phase high-performance liquid chromatography. Mass spectrometry analysis indicated the presence of a major peptide with molecular mass of approximately 5.8 kDa in both cultivars. The bactericidal activities of antimicrobial peptides were evaluated against several human pathogens that had been effective only against Klebsiella sp., a Gram-negative bacterium responsible for human urinary infection. These data indicate the biotechnological potential of sesame peptides as an alternative method for hospital infection control and also the decrease of bacterial resistance to synthetic antibiotics.