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1.
Arch Biochem Biophys ; 691: 108487, 2020 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-32710881

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/química
2.
PLoS One ; 9(3): e90487, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24614014

RESUMO

Zantedeschia aethiopica is an evergreen perennial plant cultivated worldwide and commonly used for ornamental and medicinal purposes including the treatment of bacterial infections. However, the current understanding of molecular and physiological mechanisms in this plant is limited, in comparison to other non-model plants. In order to improve understanding of the biology of this botanical species, RNA-Seq technology was used for transcriptome assembly and characterization. Following Z. aethiopica spathe tissue RNA extraction, high-throughput RNA sequencing was performed with the aim of obtaining both abundant and rare transcript data. Functional profiling based on KEGG Orthology (KO) analysis highlighted contigs that were involved predominantly in genetic information (37%) and metabolism (34%) processes. Predicted proteins involved in the plant circadian system, hormone signal transduction, secondary metabolism and basal immunity are described here. In silico screening of the transcriptome data set for antimicrobial peptide (AMP) -encoding sequences was also carried out and three lipid transfer proteins (LTP) were identified as potential AMPs involved in plant defense. Spathe predicted protein maps were drawn, and suggested that major plant efforts are expended in guaranteeing the maintenance of cell homeostasis, characterized by high investment in carbohydrate, amino acid and energy metabolism as well as in genetic information.


Assuntos
Flores/genética , Flores/metabolismo , Transcriptoma/genética , Zantedeschia/genética , Sequência de Aminoácidos , Anti-Infecciosos/farmacologia , Proteínas de Transporte/química , Ritmo Circadiano/genética , Meio Ambiente , Escherichia coli/efeitos dos fármacos , Flores/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Interações Hospedeiro-Patógeno/genética , Ligantes , Testes de Sensibilidade Microbiana , Simulação de Dinâmica Molecular , Dados de Sequência Molecular , Reguladores de Crescimento de Plantas/metabolismo , Imunidade Vegetal/efeitos dos fármacos , Imunidade Vegetal/genética , Metabolismo Secundário/efeitos dos fármacos , Metabolismo Secundário/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Staphylococcus aureus/efeitos dos fármacos , Extratos de Tecidos , Transcrição Gênica/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos , Zantedeschia/efeitos dos fármacos , Zantedeschia/imunologia
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