Translocase MraY is the site of action of lysis protein E from bacteriophage ϕX174. Previous genetic studies have shown that mutation F288L in transmembrane helix 9 of E. coli MraY confers resistance to protein E. Construction of a helical wheel model for transmembrane helix 9 of MraY and the transmembrane domain of protein E enabled the identification of an Arg-Trp-x-x-Trp (RWxxW) motif in protein E that might interact with Phe288 of MraY and the neighbouring Glu287. This motif is also found in a number of cationic antimicrobial peptide sequences. Synthetic dipeptides and pentapeptides based on the RWxxW consensus sequence showed inhibition of particulate E. coli MraY activity (IC50 200-600 μM), and demonstrated antimicrobial activity against E. coli (MIC 31-125 μg mL(-1)). Cationic antimicrobial peptides at a concentration of 100 μg mL(-1) containing Arg-Trp sequences also showed 30-60 % inhibition of E. coli MraY activity. Assay of the synthetic peptide inhibitors against recombinant MraY enzymes from Bacillus subtilis, Pseudomonas aeruginosa, and Micrococcus flavus (all of which lack Phe288) showed reduced levels of enzyme inhibition, and assay against recombinant E. coli MraY F288L and an E287A mutant demonstrated either reduced or no detectable enzyme inhibition, thus indicating that these peptides interact at this site. The MIC of Arg-Trp-octyl ester against E. coli was increased eightfold by overexpression of mraY, and was further increased by overexpression of the mraY mutant F288L, also consistent with inhibition at the RWxxW site. As this site is on the exterior face of the cytoplasmic membrane, it constitutes a potential new site for antimicrobial action, and provides a new cellular target for cationic antimicrobial peptides.

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Magnetron sputter deposition of metal targets over liquids allows producing colloidal solutions of small metal nanoparticles (NPs) without any additional reducing or stabilizing reagents. Despite that this synthetic approach is known for almost 15 years, the detailed mechanism of NP formation is still unclear. Detailed investigations must be carried out to better understand the growth mechanism and, ultimately, control the properties of the NPs. Here, the combination of the gold (Au) target and castor oil, a highly available green solvent, was chosen as a model system to investigate how different experimental parameters affect the growth of NPs. The effect of deposition time, applied sputter power, working gas pressure, and type of sputter plasma (direct current magnetron sputtering (DC-MS) vs. high-power impulse magnetron sputtering (HiPIMS)) on properties of Au NPs has been studied by UV-vis spectroscopy and transmission electron microscopy (TEM), and further supported by quantum-chemistry calculations and mass-spectrometry analysis. The mechanism of the Au NP formation includes the production of primary NPs and their subsequent aggregative growth limited by diffusion in the viscous castor oil medium. Final Au NPs have a narrow size distribution and a medium diameter of 2.4–3.2 nm when produced in DC-MS mode. The NP size can be increased up to 5.2 ± 0.8 nm by depositing in HiPIMS mode which, therefore, mimics energy and time-consuming post synthesis annealing.

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