Controlled biosilification using self-assembled short peptides A6K and V6K

Effects of external force fields on peptide self-assembly and biomimetic silica synthesis

Matrix metalloproteinases (MMPs) play critical roles in numerous physiological processes and are essential for various fundamental biological activities. Alterations in their expression levels have been associated with multiple diseases and pathological conditions. The self-assembly of small-molecule peptides into hydrogels and nanomaterials has been widely investigated for its various biomedical applications. MMPs can trigger enzyme-instructed assemblies of peptides with specific sequences to form various hydrogels for potential applications in treating several diseases, including cancer, inflammation, wound healing, myocardial infarction, and central nervous system disease, and in tissue engineering. In addition, MMPs can also trigger the self-assembly of peptides into a variety of nanostructures tailored on demand, especially serving as indicators and activators to increase their functionality in sensing applications and controlled substance release, acting as primitive biological triggers, modulating biological responses, ensuring the targeted delivery of biomolecules to specific locations, and inducing changes in the self-assembled peptide nanostructures by introducing enzyme cleavage site recognition sequences to release loaded drugs, which is crucial for applications in hydrogels and nanomaterials. This review summarizes and highlights the successful applications of various MMP-triggered self-assembled peptide hydrogels and nanostructures over the past decade and emphasizes their potential for controlled drug delivery, targeted therapy, and intelligent biomaterial-assisted wound regeneration and healing. Furthermore, future opportunities and challenges in this field are reviewed.

In nature, biological nanomaterials are synthesized under ambient conditions in a natural microscopic-sized laboratory, such as a cell. Biological molecules, such as peptides and proteins, undergo self-assembly processes in vivo and in vitro, and these monomers are assembled into various nanometer-scale structures at room temperature and atmospheric pressure. The self-assembled peptide nanostructures can be further organized to form nanowires, nanotubes, and nanoparticles via their molecular-recognition functions. The application of molecular self-assemblies of synthetic peptides as nanometer-scale building blocks in devices is robust, practical, and affordable due to their advantages of reproducibility, large-scale production ability, monodispersity, and simpler experimental methods. It is also beneficial that smart functionalities can be added at desired positions in peptide nanotubes through well-established chemical and peptide syntheses. These features of peptide-based nanotubes are the driving force for investigating and developing peptide nanotube assemblies for biological and non-biological applications.

Self-assembled peptide nanostructures have been increasingly exploited as functional materials for applications in biomedicine and energy. The emergent properties of these nanomaterials determine the applications for which they can be exploited. It has recently been appreciated that nanomaterials composed of multicomponent coassembled peptides often display unique emergent properties that have the potential to dramatically expand the functional utility of peptide-based materials. This review presents recent efforts in the development of multicomponent peptide assemblies. The discussion includes multicomponent assemblies derived from short low molecular weight peptides, peptide amphiphiles, coiled coil peptides, collagen, and β-sheet peptides. The design, structure, emergent properties, and applications for these multicomponent assemblies are presented in order to illustrate the potential of these formulations as sophisticated next-generation bio-inspired materials.

Self-assembled peptide and protein nanostructures for anti-cancer therapy: Targeted delivery, stimuli-responsive devices and immunotherapy.

We developed novel activatable probe using self-assembled peptide nanowires with low affinity and toxicity to tumor cells in the absence of matrix metalloproteinase that showed activated high affinity and toxicity and provided a highly selective and efficient platform for targeted therapy and tumor imaging.

Self-assembly of A6D amphiphilic peptides in explicit water is studied by using coarse-grained molecular dynamics simulations. It is observed that the self-assembly of randomly distributed A6D peptides leads to the formation of a network of nano-fibers. Two other simulations with cylindrical nano-fibers as the initial configuration show the dynamic stability of the self-assembled nano-fibers. As a striking feature, notable fluctuations occur along the axes of the nano-fibers. Depending on the number of peptides per unit length of the nano-fiber, flat-shaped bulges or spiral shapes along the nano-fiber axis are observed at the fluctuations. Analysis of the particle distribution around the nano-fiber indicates that the hydrophobic core and the hydrophilic shell of the nano-structure are preserved in both simulations. The size of the deformations and their correlation times are different in the two simulations. This study gives new insights into the dynamics of the self-assembled nano-structures of short amphiphilic peptides.

Sulfate ion-triggered self-assembly transitions of amphiphilic short peptides by force balance adjustment

The advancement of diverse electrochemistry technologies depends on the development of novel proton conducting polymers. Inspired by the efficacy of proton transport through proteins, we show in this work that self-assembling peptide nanostructures may be a promising alternative for such organic proton conducting materials. We demonstrate that aromatic amino acids, which participate in charge transport in nature, unprecedentedly promote proton conduction under both high and low relative humidity conditions for d,l α-cyclic peptide nanotubes. For dehydrated networks long-range order of the assemblies, induced by the aromatic side chains, is shown to be a dominating factor for promoting conductivity. However, for hydrated networks this order of effect is less significant and conductivity can be improved by the introduction of proton donating carboxylic acid peptide side chains in addition to the aromatic side chains despite the lower order of the assemblies. Based on these observations, a novel cyclic peptide that incorporates non-natural naphthyl side chains was designed. Self-assembled nanotubes of this peptide show greatly improved dehydrated conductivity, while maintaining high conductivity under hydrated conditions. We envision that the demonstrated modularity and versatility of these bio inspired nanostructures will make them extremely attractive building blocks for the fabrication of devices for energy conversion and storage applications, as well as other applications that involve proton transport, whether dry or wet conductivity is desired.

Self-assembling peptides that are capable of adopting β-sheet structures can generate nanofibers that lead to hydrogel formation. Herein, to tune the supramolecular morphologies, mechanical properties, and stimuli responses of the hydrogels, we investigated glycine substitution in a β-sheet-forming amphiphilic peptide. Glycine substitution generally enhances conformational flexibility. Indeed, glycine substitution in an amphiphilic peptide weakened the hydrogels or even inhibited the gelation. However, unexpectedly, glycine substitution at the center of the peptide molecule significantly enhanced the hydrogel stiffness. The central glycine substitution affected the molecular packing and led to twisted β-sheet structures and to nanofiber bundling, which likely led to the stiffened hydrogel. Importantly, the supramolecular structures were accurately predicted by molecular dynamics simulations, demonstrating the helpfulness of these techniques for the identification of self-assembling peptides. The hydrogel formed by the amphiphilic peptide with the central glycine substitution had cell adhesive function, and showed a reversible thermal gel-to-sol transition. Thus, glycine substitution is effective in modulating self-assembling structures, rheological properties, and dynamics of biofunctional self-assembling peptides.

In this study, a modified dehydropeptide, Boc-FΔF-εAhx-OH, was conjugated with an aminoglycoside antibiotic, neomycin, to construct a multifunctional conjugate, Pep-Neo. The amphiphilic conjugate (Pep-Neo) was able to self-assemble into cationic nanostructures in an aqueous solution at low concentrations. Nanostructure formation was evidenced by TEM and dynamic light scattering analyses. The average hydrodynamic diameter of the self-assembled Pep-Neo nanostructures was found to be ∼279 nm with a zeta potential of +28 mV. The formation of nanostructures with a hydrophobic core and cationic hydrophilic shell resulted in an increased local concentration of cationic charge (ca. in 50% aqueous methanol, i.e. disassembled structure, zeta potential decreased to +17.6 mV), leading to efficient interactions with negatively charged plasmid DNA (pDNA). The size and zeta potential of the resulting Pep-Neo/pDNA complex were found to be ∼154 nm and +19.4 mV, respectively. Having been characterized by physicochemical techniques, the complex was evaluated for its toxicity and ability to deliver nucleic acid therapeutics. The flow cytometry results on MCF-7 cells revealed that Pep-Neo/pDNA complex transfected ∼27% cells at a w/w ratio of 66.6 while the standard transfection reagent, Lipofectamine, could transfect only ∼15% cells. MTT and hemolysis assays showed the non-toxic nature of the projected conjugate at various concentrations. Further, these nanostructures were shown to encapsulate hydrophobic drugs in the core. Finally, Pep-Neo nanostructures showed efficient antibacterial activity against different strains of Gram-positive and -negative bacteria. Interestingly, unlike neomycin, which is highly effective against Gram-negative bacteria, these nanostructures showed considerably high efficiency against Gram-positive strains, highlighting the promising potential of these nanostructures for various biomedical applications.

The development of modern nanomedicine greatly depends on the involvement of novel materials as drug delivery system. In order to maximize the therapeutic effects of drugs and minimize their side effects, a number of natural or synthetic materials have been widely investigated for drug delivery. Among these materials, biomimetic self-assembling peptides (SAPs) have received more attention in recent years. Considering the rapidly growing number of SAPs designed for drug delivery, a summary of how SAPs-based drug delivery systems were designed, would be beneficial. We outlined research works on different SAPs that have been investigated as carriers for different drugs, focusing on the design of SAPs nanomaterials and how they were used for drug delivery in different strategies. Based on the principle rules of chemical complementarity and structural compatibility, SAPs such as ionic self-complementary peptide, peptide amphiphile and surfactant-like peptide could be designed. Determined by the features of peptide materials and the drugs to be delivered, different strategies such as hydrogel embedding, hydrophobic interaction, electrostatic interaction, covalent conjugation or the combination of them could be employed to fabricate SAPs-drug complex, which could achieve slow release, targeted or environment-responsive delivery of drugs. Furthermore, some SAPs could also be combined with other types of materials for drug delivery, or even act as drug by themselves. Various types of SAPs have been designed and used for drug delivery following various strategies, suggesting that SAPs as a category of versatile nanomaterials have promising potential in the field of nanomedicine.

Protein nanowires exhibiting specific biological activities hold promise for interacting with living cells and controlling and predicting biological responses such as apoptosis, endocytosis, and cell adhesion. In this chapter, fabrication of various size-controlled protein nanowires by single-particle nanofabrication technique (SPNT) is reviewed. The human serum albumin (HSA) nanowires of various lengths can be fabricated, from HSA nanodots to HSA nanowires with extremely high aspect ratios of over 1000, by controlling the thickness of the film. The diameters of the protein nanowires are almost equal to the diameter of self-assembled protein and peptide nanostructures. Degradation of the HSA nanowires was examined using protease. The biotinylated human serum albumin nanowires can be transformed into nanowires exhibiting a biological function such as an enzyme through using avidin–biotin interaction. Interestingly, 2D Protein sheets and protein synthetic polymer nanowires can be fabricated by the application of SPNT. SPNT is a versatile platform for functionalizing the surface of any protein molecule with an extremely large surface area.

Essential amino acids in catalytic sites of native enzymes are important in nature inspired catalyst designs. Active sites of enzymes contain the coordinated assembly of multiple amino acids, and catalytic action is generated by the dynamic interactions among multiple residues. However, catalysis studies are limited by the complex and dynamic structure of the enzyme; and it is difficult to exclusively attribute a given function to a specific residue. Minimalistic approaches involving artificial catalytic sites are promising for the investigation of the enzyme function in the absence of non-essential protein components, and self-assembling peptide nanostructures are especially advantageous in this context. Here we demonstrate the design and characterization of an enzyme-mimetic catalytic nanosystem presenting essential residues (Ser, His, Asp). The function of each residue and its combinations on the nanostructures in hydrolysis reaction was studied. The catalytic self-assembled nanostructures were used for efficient ester hydrolysis such as a model substrate (pNPA) and a natural substrate (acetylcholine) highlighting the key role of self-assembly in catalytic domain formation to test the efficiency of the de novo designed catalyst as a catalytic triad model.

Supramolecular self-assembled bio-inspired peptide nanostructures are favorable to be implemented in diverse nanophotonics applications due to their superior physical properties such as wideband optical transparency, high second-order nonlinear response, waveguiding properties and more. Here, we focus on the optical properties found in di-phenylalanine peptide nano-architectures, with special emphasize on their linear and nonlinear optical waveguiding effects. Using both simulation and experiments, we show their ability to passively guide light at both fundamental and second-harmonic frequencies. In addition, we show that at elevated temperatures, 140-180°C, these native supramolecular structures undergo irreversible thermally induced transformation via re-assembling into completely new thermodynamically stable phase having nanofiber morphology similar to those of amyloid fibrils. In this new phase, the peptide nanofibers lose their second-order nonlinear response, while exhibit profound modification of optoelectronic properties followed by the appearance of visible (blue and green) photoluminescence (PL). Our study propose a new generation of multifunctional optical waveguides with variety of characteristics, which self-assembled into 1D-elongated nanostructures and could be used as building blocks of many integrated photonic devices.