RESUMEN
Protein (mis)folding, stability and aggregation are of interest in numerous fields, such as food sciences, biotechnology, and health sciences, and efforts are directed towards the elucidation of the underlying molecular mechanisms. Through an integrative approach, we show that a subtle balance between hydrogen bond formation and hydrophobic interactions defines protein self-assembly pathways. Hydrophobic co-solvents, such as monohydric alcohols, modulate these two forces through a combination of direct solvent-protein and solvent-mediated interactions, depending on the size of the alcohol. This affects the initial conformation of the model protein α-lactalbumin, which can be linked to variations of its fibrillation propensity, as well as the morphology of the final structures. These findings pave the way towards a better understanding of the forces governing protein self-assembly, allowing the development of strategies to suppress unwanted aggregation and control the growth of tuneable protein-based biomaterials.
Asunto(s)
Lactalbúmina/química , Conformación Proteica , Pliegue de Proteína , Multimerización de Proteína , Solventes/química , Alcoholes/química , Animales , Bovinos , Enlace de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Simulación de Dinámica MolecularRESUMEN
Gram-negative bacterial pathogens are intrinsically resistant to several antibiotics that are not able to penetrate the cell envelope barrier. The aim of this study was to identify peptides that at low concentrations induce susceptibility to these antibiotics in multidrug-resistant (MDR) Gram-negative bacterial strains of clinical relevance. Pairwise screening of 34 diverse peptides and four antibiotics (erythromycin, linezolid, rifampicin and vancomycin) with primary activity against Gram-positive bacteria identified 4 peptides that at submicromolar concentrations conferred susceptibility to rifampicin or erythromycin in Escherichia coli ATCC 25922. The identified peptides exhibited synergy with azithromycin and potentiated clindamycin in MDR E. coli ST131 and Klebsiella pneumoniae ST258. The low cytotoxicity toward eukaryotic cells (IC50 > 50 µM) observed for two of these peptides (KLWKKWKKWLK-NH2 and GKWKKILGKLIR-NH2) prompted synthesis and evaluation of the corresponding all-d analogues (D1 and D2), which retained similar synergistic antibacterial profiles. Low concentrations of D1 and D2 in combination with azithromycin and rifampicin inhibited growth of most clinical E. coli, K. pneumoniae and Acinetobacter baumannii strains tested. These data demonstrate that combinatorial screening at low peptide concentrations constitutes an efficient approach to identify clinically relevant peptide-antibiotic combinations. In vivo pharmacokinetic/pharmacodynamic and toxicity studies are needed to further validate the use of the peptides identified in this study for repurposing azithromycin and rifampicin against Gram-negative pathogens.
Asunto(s)
Antibacterianos/farmacología , Azitromicina/farmacología , Reposicionamiento de Medicamentos , Sinergismo Farmacológico , Bacterias Gramnegativas/efectos de los fármacos , Péptidos/farmacología , Rifampin/farmacología , Acinetobacter baumannii , Supervivencia Celular/efectos de los fármacos , Escherichia coli , Células Eucariotas/efectos de los fármacos , Humanos , Concentración 50 Inhibidora , Klebsiella pneumoniae , Péptidos/toxicidadRESUMEN
Infections with multidrug-resistant pathogens are an increasing concern for public health. Recently, subtypes of peptide-peptoid hybrids were demonstrated to display potent activity against multidrug-resistant Gram-negative bacteria. Here, structural variation of these antibacterial peptidomimetics was investigated as a tool for optimizing cell selectivity. A protocol based on dimeric building blocks allowed for efficient synthesis of an array of peptide-peptoid oligomers representing length variation as well as different backbone designs displaying chiral or achiral peptoid residues. Lack of α-chirality in the side chains of the peptoid residues proved to be correlated to reduced cytotoxicity. Furthermore, optimization of the length of these peptidomimetics with an alternating cationic-hydrophobic design was a powerful tool to enhance the selectivity against Gram-negative pathogens over benign mammalian cells. Thus, lead compounds with a high selectivity toward killing of clinically important multidrug-resistant E. coli were identified.