Lactoferricin-Derived L5a Cell-Penetrating Peptide for Delivery of DNA into Cells

Cell-penetrating peptides (CPPs) are small peptides which help intracellular delivery of functional macromolecules, including DNAs, RNAs, and proteins, across the cell membrane and into the cytosol, and even into the nucleus in some cases. Delivery of macromolecules can facilitate transfection, aid in gene therapy and transgenesis, and alter gene expression. L5a (RRWQW), originally derived from bovine lactoferricin, is one kind of CPPs which can promote cellular uptake of plasmid DNA and enters cells via direct membrane translocation. The peptide complexes noncovalently with DNA over a short incubation period. DNA plasmid and L5a complex stability is confirmed by a decrease in mobility in a gel retardation assay, and successful transfection is proven by the detection of a reporter gene in cells using fluorescent microscopy. Here, we describe methods to study noncovalent interactions between L5a and plasmid DNA, and the delivery of L5a/DNA complexes into cells. L5a is the one of the smallest CPPs discovered to date, providing a small delivery vehicle for macromolecules in mammalian cells. A small vehicle which can enter the nucleus is ideal for efficient gene uptake, transfer, and therapy. It is simple to complex with DNA plasmids, and its nature allows mammalian cells to be easily transfected

Antimicrobial and Cell-Penetrating Peptides: How to Understand Two Distinct Functions Despite Similar Physicochemical Properties

Antimicrobial and cell-penetrating peptides are both classes of membrane-active peptides sharing similar physicochemical properties. Both kinds of peptides have attracted much attention owing to their specific features. AMPs disrupt cell membranes of bacteria and display urgently needed antibiotic substances with alternative modes of action. Since the multidrug resistance of bacterial pathogens is a more and more raising concern, AMPs have gained much interest during the past years. On the other side, CPPs enter eukaryotic cells without substantially affecting the plasma membrane. They can be used as drug delivery platforms and have proven their usefulness in various applications. However, although both groups of peptides are quite similar, their intrinsic activity is often different, and responsible factors are still in discussion. The aim of this chapter is to summarize and shed light on recent findings and concepts dealing with differences and similarities of AMPs and CPPs and to understand these different functions

Exploring the Sequence Space for (tri-) Peptide Self-assembly to Design and Discover New Hydrogels

NoPeptides that self-assemble into nanostructures are of tremendous interest for biological, medical, photonic and nanotechnological applications. The enormous sequence space that is available from 20 amino acids probably harbours many interesting candidates, but it is currently not possible to predict supramolecular behaviour from sequence alone. Here, we demonstrate computational tools to screen for the aqueous self-assembly propensity in all of the 8,000 possible tripeptides and evaluate these by comparison with known examples. We applied filters to select for candidates that simultaneously optimize the apparently contradicting requirements of aggregation propensity and hydrophilicity, which resulted in a set of design rules for self-assembling sequences. A number of peptides were subsequently synthesized and characterized, including the first reported tripeptides that are able to form a hydrogel at neutral pH. These tools, which enable the peptide sequence space to be searched for supramolecular properties, enable minimalistic peptide nanotechnology to deliver on its promise