Structural insights into and activity analysis of the antimicrobial peptide myxinidin.

The marine environment has been poorly explored in terms of potential new molecules possessing antibacterial activity. Antimicrobial peptides (AMPs) offer a new potential class of pharmaceuticals; however, further optimization is needed if AMPs are to find broad use as antibiotics. We focused our studies on a peptide derived from the epidermal mucus of hagfish (Myxine glutinosa L.), which was previously characterized and showed high antimicrobial activity against human and fish pathogens. In the present work, the activities of myxinidin peptide analogues were analyzed with the aim of widening the original spectrum of action of myxinidin by suitable changes in the peptide primary structure. The analysis of key residues by alanine scanning allowed for the design of novel peptides with increased activity. We identified the amino acids that are of the utmost importance for the observed antimicrobial activities against a set of pathogens comprising both Gram-negative and Gram-positive bacteria. Overall, optimized bactericidal potency was achieved by adding a tryptophan residue at the N terminus and by the simultaneous substitution of residues present in positions 3, 4, and 11 with arginine. These results indicate that the myxinidin analogues emerge as an attractive alternative for treating drug-resistant infectious diseases and provide key insights into a rational design for novel agents against these pathogens.

Structure-activity relations of myxinidin, an antibacterial peptide derived from the epidermal mucus of hagfish.

The structure-activity relations of myxinidin, a peptide derived from epidermal mucus of hagfish, Myxine glutinosa L., were investigated. Analysis of key residues allowed us to design new peptides with increased efficiency. Antimicrobial activity of native and modified peptides demonstrated the key role of uncharged residues in the sequence; the loss of these residues reduces almost entirely myxinidin antimicrobial activity, while insertion of arginine at charged and uncharged position increases antimicrobial activity compared with that of native myxinidin. Particularly, we designed a peptide capable of achieving a high inhibitory effect on bacterial growth. Experiments were conducted using both Gram-negative and Gram-positive bacteria. Nuclear magnetic resonance (NMR) studies showed that myxinidin is able to form an amphipathic α-helical structure at the N terminus and a random coil region at the C terminus.

A peptide derived from herpes simplex virus type 1 glycoprotein H: membrane translocation and applications to the delivery of quantum dots.

Cell membranes are impermeable to most molecules that are not actively imported by living cells, including all macromolecules and even small molecules whose physiochemical properties prevent passive membrane diffusion. However, recently, we have seen the development of increasingly sophisticated methodology for intracellular drug delivery. Cell-penetrating peptides (CPPs), short peptides believed to enter cells by penetrating cell membranes, have attracted great interest in the hope of enhancing gene therapy, vaccine development and drug delivery. Nevertheless, to achieve an efficient intracellular delivery, further strategies to bypass the endocytotic pathway must be investigated. We report on a novel peptide molecule derived from glycoprotein gH of herpes simplex type I virus that is able to traverse the membrane bilayer and to transport a cargo into the cytoplasm with novel properties in comparison with existing CPPs. We use as cargo molecule quantum dots that do not significantly traverse the membrane bilayer on their own. FROM THE CLINICAL EDITOR: Cell-penetrating peptides have recently attracted great interest in optimizing gene therapy, vaccine development and drug delivery. In this study, a peptide derived from glycoprotein gH of herpes simplex I is investigated from this standpoint.

Pectin functionalized with natural fatty acids as antimicrobial agent.

Several pectin derivatives were prepared by chemical modifications of the polysaccharide with natural fatty acids. The obtained biodegradable pectin-based materials, pectin-linoleate, pectin-oleate and pectin-palmitate, were investigated for their antimicrobial activity against several bacterial strains, Staphylococcus aureus and Escherichia coli. Good results were obtained for pectin-oleate and pectin-linoleate, which inhibit the growth of the selected microorganisms by 50-70%. They exert the better antimicrobial activity against S. aureus. Subsequently, the pectin-oleate and the pectin-linoleate samples were coated on polyethylene films and were assessed for their capacity to capture the oxygen molecules, reducing its penetration into the polymeric support. These results confirmed a possible application of the new materials in the field of active food packaging.

Cellular cholesterol involvement in Src, PKC, and p38/JNK transduction pathways by porins.

Biological membranes are described as a mosaic of different domains where interactions between membrane components induce the formation of subdomains with different characteristics and functions. Lipids play an important role in the formation of lipid-enriched microdomains where they dynamically associate to form platforms important for membrane protein sorting and construction of signaling complexes. Cholesterol confined in lipid domains is a crucial component required by microorganisms, directly or indirectly, to enter or exit the intracellular compartment. Cellular activation mediated by superficial bacterial component may be modified by local cholesterol depletion. Therefore, new perspectives for unconventional therapeutic intervention in Gram-negative infections may be envisaged. We tested this hypothesis by using methyl-beta-cyclodextrin (mbetaCD) as a cholesterol-complexing agent to alter the U937 plasma membrane cholesterol content. Our results demonstrate that cholesterol depletion of U937 cells inhibited Salmonella enterica serovar Typhimurium porins-mediated phosphorylation of Src kinase family, protein kinase C (PKC), JNK, and p38, while cholesterol repletion restored the phosphorylation. Lipopolysaccharide (LPS) extracted from the same bacterial strain has been used as a control. Our data demonstrate that the lack of activation of signal transduction pathway observed following cholesterol depletion differently modulates the release of interleukin-6 (IL-6) or tumor necrosis factor-alpha (TNF-alpha), suggesting that Src, associated to lipid domains, may represent an important pathway in Gram-negative-induced cellular signal.