RESUMO
Aquaporins (AQPs) are small transmembrane tetrameric proteins that facilitate water, solute and gas exchange. Their presence has been extensively reported in the biological membranes of almost all living organisms. Although their discovery is much more recent than ion transport systems, different biophysical approaches have contributed to confirm that permeation through each monomer is consistent with closed and open states, introducing the term gating mechanism into the field. The study of AQPs in their native membrane or overexpressed in heterologous systems have experimentally demonstrated that water membrane permeability can be reversibly modified in response to specific modulators. For some regulation mechanisms, such as pH changes, evidence for gating is also supported by high-resolution structures of the water channel in different configurations as well as molecular dynamics simulation. Both experimental and simulation approaches sustain that the rearrangement of conserved residues contributes to occlude the cavity of the channel restricting water permeation. Interestingly, specific charged and conserved residues are present in the environment of the pore and, thus, the tetrameric structure can be subjected to alter the positions of these charges to sustain gating. Thus, is it possible to explore whether the displacement of these charges (gating current) leads to conformational changes? To our knowledge, this question has not yet been addressed at all. In this review, we intend to analyze the suitability of this proposal for the first time.
Assuntos
Aquaporinas , Aquaporinas/metabolismo , Simulação de Dinâmica Molecular , Água/metabolismo , Biofísica , Permeabilidade da Membrana CelularRESUMO
Host defense peptides (HDPs) exhibit a broad range of antimicrobial and immunomodulatory activities. In this sense, both functions are like different sides of the same coin. The direct antimicrobial side was discovered first, and widely studied for the development of anti-infective therapies. In contrast, the immunomodulatory side was recognized later and in the last 20 years the interest in this field has been continuously growing. Different to their antimicrobial activities, the immunomodulatory activities of host defense peptides are more effective in vivo. They offer a great opportunity for new therapeutic applications in the fields of anti-infective therapy, chronic inflammatory diseases treatment, novel vaccine adjuvants development and anticancer immunotherapy. These immune related functions of HDPs includes chemoattraction of leukocytes, modulation of inflammation, enhancement of antigen presentation and polarization of adaptive immune responses. Our attempt with this review is to make a careful evaluation of different aspects of the less explored, but attractive immunomodulatory side of the HDP functional coin.
Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Fatores Imunológicos/farmacologia , Animais , Anti-Infecciosos/farmacologia , Humanos , Sistema Imunitário/efeitos dos fármacosRESUMO
Cm-p5 is a snail-derived antimicrobial peptide, which demonstrated antifungal activity against the pathogenic strains of Candida albicans. Previously we synthetized a cyclic monomer as well as a parallel and an antiparallel dimer of Cm-p5 with improved antifungal activity. Considering the alarming increase of microbial resistance to conventional antibiotics, here we evaluated the antimicrobial activity of these derivatives against multiresistant and problematic bacteria and against important viral agents. The three peptides showed a moderate activity against Pseudomonas aeruginosa, Klebsiella pneumoniae Extended Spectrum ß-Lactamase (ESBL), and Streptococcus agalactiae, with MIC values > 100 µg/mL. They exerted a considerable activity with MIC values between 25-50 µg/mL against Acinetobacter baumanii and Enterococcus faecium. In addition, the two dimers showed a moderate activity against Pseudomonas aeruginosa PA14. The three Cm-p5 derivatives inhibited a virulent extracellular strain of Mycobacterium tuberculosis, in a dose-dependent manner. Moreover, they inhibited Herpes Simplex Virus 2 (HSV-2) infection in a concentration-dependent manner, but had no effect on infection by the Zika Virus (ZIKV) or pseudoparticles of Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2). At concentrations of >100 µg/mL, the three new Cm-p5 derivatives showed toxicity on different eukaryotic cells tested. Considering a certain cell toxicity but a potential interesting activity against the multiresistant strains of bacteria and HSV-2, our compounds require future structural optimization.
Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Antivirais/farmacologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Herpesvirus Humano 2/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Antibacterianos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Antivirais/química , Candida albicans/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Dimerização , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , SARS-CoV-2/efeitos dos fármacosRESUMO
Antimicrobial peptides (AMPs) are biomolecules with antimicrobial activity against a broad group of pathogens. In the past few decades, AMPs have represented an important alternative for the treatment of infectious diseases. Their isolation from natural sources has been widely investigated. In this sense, mollusks are promising organisms for the identification of AMPs given that their immune system mainly relies on innate response. In this report, we characterized the peptide fraction of the Cuban freshwater snail Pomacea poeyana (Pilsbry, 1927) and identified 37 different peptides by nanoLC-ESI-MS-MS technology. From these peptide sequences, using bioinformatic prediction tools, we discovered two potential antimicrobial peptides named Pom-1 (KCAGSIAWAIGSGLFGGAKLIKIKKYIAELGGLQ) and Pom-2 (KEIERAGQRIRDAIISAAPAVETLAQAQKIIKGG). Database search revealed that Pom-1 is a fragment of Closticin 574 previously isolated from the bacteria Clostridium tyrobutyrium, and Pom-2 is a fragment of cecropin D-like peptide first isolated from Galleria mellonella hemolymph. These sequences were chemically synthesized and evaluated against different human pathogens. Interestingly, structural predictions of both peptides in the presence of micelles showed models that comprise two alpha helices joined by a short loop. The CD spectra analysis of Pom-1 and Pom-2 in water showed for both structures a high random coil content, a certain content of α-helix and a low ß-sheet content. Like other described AMPs displaying a disordered structure in water, the peptides may adopt a helical conformation in presence of bacterial membranes. In antimicrobial assays, Pom-1 demonstrated high activity against the Gram-negative bacteria Pseudomonas aeruginosa and moderate activity against Klebsiella pneumoniae and Listeria monocytogenes. Neither of the two peptides showed antifungal action. Pom-1 moderately inhibits Zika Virus infection but slightly enhances HIV-1 infectivion in vitro. The evaluation of cell toxicity on primary human macrophages did not show toxicity on THP-1 cells, although slight overall toxicity was observed in high concentrations of Pom-1. We assume that both peptides may play a key role in innate defense of P. poeyana and represent promising antimicrobial candidates for humans.
Assuntos
Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Antivirais/farmacologia , Klebsiella pneumoniae/efeitos dos fármacos , Listeria monocytogenes/efeitos dos fármacos , Moluscos/química , Animais , Antibacterianos/química , Peptídeos Catiônicos Antimicrobianos/química , Antivirais/química , Sobrevivência Celular/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Células THP-1 , Infecção por Zika virus/tratamento farmacológicoRESUMO
Following the information obtained by a rational design study, a cyclic and helical-stabilized analogue of the peptide Cm-p5 was synthetized. The cyclic monomer showed an increased activity in vitro against Candida albicans and Candida parapsilosis, compared to Cm-p5. Initially, 14 mutants of Cm-p5 were synthesized following a rational design to improve the antifungal activity and pharmacological properties. Antimicrobial testing showed that the activity was lost in each of these 14 analogues, suggesting, as a main conclusion, that a Glu-His salt bridge could stabilize Cm-p5 helical conformation during the interaction with the plasma membrane. A derivative, obtained by substitution of Glu and His for Cys, was synthesized and oxidized with the generation of a cyclic monomer with improved antifungal activity. In addition, two dimers were generated during the oxidation procedure, a parallel and antiparallel one. The dimers showed a helical secondary structure in water, whereas the cyclic monomer only showed this conformation in SDS. Molecular dynamic simulations confirmed the helical stabilizations for all of them, therefore indicating the possible essential role of the Glu-His salt bridge. In addition, the antiparallel dimer showed a moderate activity against Pseudomonas aeruginosa and a significant activity against Listeria monocytogenes. Neither the cyclic monomer nor the dimers were toxic against macrophages or THP-1 human cells. Due to its increased capacity for fungal control compared to fluconazole, its low cytotoxicity, together with a stabilized α-helix and disulfide bridges, that may advance its metabolic stability, and in vivo activity, the new cyclic Cm-p5 monomer represents a potential systemic antifungal therapeutic candidate.
RESUMO
Antimicrobial peptides are small cationic molecules present in almost all living organisms. They show direct or indirect (immunomodulation) activity in a wide range of pathogenic microorganisms as members of the humoral arsenal of innate immunity. In mammals they play a significant role in respiratory airways. The most abundant antimicrobial peptides in the respiratory tract of mammals are lysozymes, lactoferrin, histatins, defensins and cathelicidins. Respiratory and pulmonary infections are combated, primarily, by antimicrobial peptides like LL-37 against Gram-negative bacteria, histatin 5 against Candida albicans and human peptides from neutrophils against adenovirus, influenza and parainfluenza. This paper provides a review of the most important antimicrobial peptides in the respiratory tract and their use in the search for new effective agents against microorganisms that cause respiratory infections based on information published in MedLine, the Web of Science and Scopus in recent years(AU)
Los péptidos antimicrobianos son pequeñas moléculas catiónicas presentes en casi todos los organismos vivos. Muestran actividad directa o indirecta (inmunomodulación) en una amplia gama de microorganismos patógenos como miembros del arsenal humoral de la inmunidad innata. En los mamíferos juegan un papel importante en las vías respiratorias. Los péptidos antimicrobianos más abundantes en el tracto respiratorio son lisozima, lactoferrina, histatinas, defensinas y catelicidinas. Las infecciones respiratorias y pulmonares son combatidas, principalmente, por péptidos antimicrobianos como LL-37 contra bacterias gramnegativas, histatina 5 contra Candida albicans y péptidos humanos de neutrófilos contra adenovirus, influenza y parainfluenza. Este artículo proporciona una revisión sobre los péptidos antimicrobianos más importantes en el tracto respiratorio y su empleo en la búsqueda de nuevos agentes eficaces contra microorganismos causantes de infecciones respiratorias teniendo en cuenta la información publicada al respecto en MedLine, Web of Science y Scopus en los últimos años(AU)
Assuntos
Humanos , Masculino , Feminino , Resistência Microbiana a Medicamentos , Infecções por Paramyxoviridae , Peptídeos Catiônicos Antimicrobianos/uso terapêuticoRESUMO
In the last years, the antimicrobial resistance against antibiotics has become a serious health issue, arise as global threat. This has generated a search for new strategies in the progress of new antimicrobial therapies. In this context, different nanosystems with antimicrobial properties have been studied. Specifically, magnetic nanoparticles seem to be very attractive due to their relatively simple synthesis, intrinsic antimicrobial activity, low toxicity and high versatility. Iron oxide NPs (IONPs) was authorized by the World Health Organization for human used in biomedical applications such as in vivo drug delivery systems, magnetic guided therapy and contrast agent for magnetic resonance imaging have been widely documented. Furthermore, the antimicrobial activity of different magnetic nanoparticles has recently been demonstrated. This review elucidates the recent progress of IONPs in drug delivery systems and focuses on the treatment of infectious diseases and target the possible detrimental biological effects and associated safety issues.
Assuntos
Anti-Infecciosos/administração & dosagem , Sistemas de Liberação de Medicamentos , Nanopartículas de Magnetita , Animais , Anti-Infecciosos/farmacologia , Doenças Transmissíveis/tratamento farmacológico , Doenças Transmissíveis/microbiologia , Resistência Microbiana a Medicamentos , Compostos Férricos/química , HumanosRESUMO
El aumento en la incidencia de las enfermedades infecciosas en los últimos años se ha favorecido por diferentes causas. Entre estas se destacan las inmunodeficiencias adquiridas (sida, trasplantes de órganos, quimioterapia oncológica), la migración de personas que trae consigo la posibilidad de importar enfermedades hacia poblaciones susceptibles, así como el excesivo empleo de antibióticos. Debido a esta situación se ha incrementado la búsqueda de nuevos candidatos terapéuticos para el desarrollo de terapias más efectivas. En este sentido los péptidos antimicrobianos constituyen una opción promisoria, pues presentan un amplio espectro de actividad frente a varios microorganismos patógenos. Además, se encuentran ampliamente distribuidos en la naturaleza, desde organismos unicelulares hasta mamíferos. Algunos péptidos antimicrobianos ya están siendo evaluados en estudios clínicos aunque muchos de ellos no han tenido resultados favorables in vivo debido a su poca estabilidad metabólica y toxicidad, entre otros. Con el fin de optimizar estas propiedades de los péptidos antimicrobianos se han trazado diferentes estrategias como la modificación química de su estructura y la conjugación con nanopartículas magnéticas. Es por eso que este artículo tiene el objetivo de revisar las potenciales aplicaciones terapéuticas de estas moléculas, teniendo en cuenta la información publicada al respecto en MedLine, Web of Science y Scopus en los últimos años
The growing incidence of infectious disease in recent years may be attributed to several causes, among them acquired immunodeficiencies (AIDS, organ transplant, oncological chemotherapy), human migration and the consequent import of diseases into susceptible populations, and the excessive use of antibiotics. This situation has fostered the search for new therapeutic candidates for the development of more effective treatments. Antimicrobial peptides are a promising alternative in this respect, due to their broad spectrum of activity against several pathogenic microorganisms. Moreover, they are widely distributed in nature, from unicellular organisms to mammals. Some antimicrobial peptides are already being evaluated in clinical studies, though many of them have not produced any favorable results in vivo due to their low metabolic stability and their toxicity, among other factors. Several strategies have been developed to overcome the above mentioned drawbacks, among them conjugation of microbial peptides with magnetic nanoparticles and chemical modification of their structure. The present study is aimed at reviewing the potential therapeutic applications of these molecules based on information published in MedLine, the Web of Science and Scopus in recent years.
RESUMO
Recently, several peptides have been studied regarding the defence process against pathogenic microorganisms, which are able to act against different targets, with the purpose of developing novel bioactive compounds. The present work focuses on the structural and functional evaluation of the palindromic antimicrobial peptide Pa-MAP2, designed based on the peptide Pa-MAP from Pleuronectes americanus. For a better structural understanding, molecular modelling analyses were carried out, together with molecular dynamics and circular dichroism, in different media. Antibacterial activity against Gram-negative and positive bacteria was evaluated, as well as cytotoxicity against human erythrocytes, RAW 264.7, Vero and L6 cells. In silico docking experiments, lipid vesicle studies, and atomic force microscopy (AFM) imaging were carried out to explore the activity of the peptide. In vivo studies on infected mice were also done. The palindromic primary sequence favoured an α-helix structure that was pH dependent, only present on alkaline environment, with dynamic N- and C-terminals that are stabilized in anionic media. Pa-MAP2 only showed activity against Gram-negative bacteria, with a MIC of 3.2 µM, and without any cytotoxic effect. In silico, lipid vesicles and AFM studies confirm the preference for anionic lipids (POPG, POPS, DPPE, DPPG and LPS), with the positively charged lysine residues being essential for the initial electrostatic interaction. In vivo studies showed that Pa-MAP2 increases to 100% the survival rate of mice infected with Escherichia coli. Data here reported indicated that palindromic Pa-MAP2 could be an alternative candidate for use in therapeutics against Gram-negative bacterial infections.