Technique for determination of gold electrode area and its application of characterization in self-assembling process

Underpotential deposition(UPD) of copper at polycrystal gold surface under different concentrations has been studied, and its reversibility and stability in high concentration (0.2M CuSO4+0.IM H2SO4) have been demonstrated by cyclic voltermmetry and EQCM. A Valid approach to determine the gold electrode area in presence of adsorbed species has been provided by using Cu UPD method. Further, the growth kinetics of decane thiol on gold has also been investigated based on such a Cu UPD technique.

Development and characterization of self-assembling nanoparticles using a bio-inspired amphipathic peptide for gene delivery

The design of a non-viral gene delivery vehicle capable of delivering and releasing a functional nucleic acid cargo intracellularly remains a formidable challenge. For systemic gene therapy to be successful a delivery vehicle is required that protects the nucleic acid cargo from enzymatic degradation, extravasates from the vasculature, traverses the cell membrane, disrupts the endosomal vesicles and unloads the cargo at its destination site, namely the nucleus for the purposes of gene delivery. This manuscript reports the extensive investigation of a novel amphipathic peptide composed of repeating RALA units capable of overcoming the biological barriers to gene delivery both in vitro and in vivo. Our data demonstrates the spontaneous self-assembly of cationic DNA-loaded nanoparticles when the peptide is complexed with pDNA. Nanoparticles were < 100 nm, were stable in the presence of serum and were fusogenic in nature, with increased peptide α-helicity at a lower pH. Nanoparticles proved to be non-cytotoxic, readily traversed the plasma membrane of both cancer and fibroblast cell lines and elicited reporter-gene expression following intravenous delivery in vivo. The results of this study indicate that RALA presents an exciting delivery platform for the systemic delivery of nucleic acid therapeutics.

BslA is a self-assembling bacterial hydrophobin that coats the Bacillus subtilis biofilm

Biofilms represent the predominant mode of microbial growth in the natural environment. Bacillus subtilis is a ubiquitous Gram-positive soil bacterium that functions as an effective plant growth-promoting agent. The biofilm matrix is composed of an exopolysaccharide and an amyloid fiber-forming protein, TasA, and assembles with the aid of a small secreted protein, BslA. Here we show that natively synthesized and secreted BslA forms surface layers around the biofilm. Biophysical analysis demonstrates that BslA can self-assemble at interfaces, forming an elastic film. Molecular function is revealed from analysis of the crystal structure of BslA, which consists of an Ig-type fold with the addition of an unusual, extremely hydrophobic "cap" region. A combination of in vivo biofilm formation and in vitro biophysical analysis demonstrates that the central hydrophobic residues of the cap are essential to allow a hydrophobic, nonwetting biofilm to form as they control the surface activity of the BslA protein. The hydrophobic cap exhibits physiochemical properties remarkably similar to the hydrophobic surface found in fungal hydrophobins; thus, BslA is a structurally defined bacterial hydrophobin. We suggest that biofilms formed by other species of bacteria may have evolved similar mechanisms to provide protection to the resident bacterial community.

Ultrashort self-assembling peptidomimetic nanomaterials target resistant pathogenic infections

The impending and increasing threat of antimicrobial resistance has led to a greater focus into developing alternative therapies as substitutes for traditional antibiotics for the treatment of multi-drug resistant infections.1 Our group has developed a library of short, cost-effective, diphenylalanine-based peptides (X1-FF-X2) which selective eradicate (viability reduced >90% in 24 hours) the most resistant biofilm forms of a range of Gram-positive and negative pathogens including: methicillin resistant and sensitive Staphyloccoccus aureus and Staphyloccoccus epidermidis; Pseudomonas aeruginosa, Proteus mirabilis and Escherichia coli. They demonstrate a reduced cell cytotoxic profile (NCTC929 murine fibroblast) and limited haemolysis.2 Our molecules have the ability respond to subtle changes in pH, associated with bacterial infection, self-assembling to form β-sheet secondary structures and supramolecular hydrogels at low concentrations (~0.5%w/v). Conjugation of variety of aromatic-based drugs at the X1 position, including non-steroidal anti-inflammatories (NSAIDs), confer further pharmacological properties to the peptide motif enhancing their therapeutic potential. In vivo studies using waxworms (Galleria mellonella) provide promising preliminary results demonstrating the low toxicity and high antimicrobial activity of these low molecular weight gelators in animal models. This work shows biofunctional peptide-based nanomaterials hold great promise for future translation to patients as antimicrobial drug delivery and biomaterial platforms.3 [1] G. Laverty, S.P. Gorman and B.F. Gilmore. Int.J.Mol.Sci. 2011, 12, 6566-6596. [2] G. Laverty, A.P. McCloskey, B.F. Gilmore, D.S. Jones, J Zhou, B Xu. Biomacromolecules. 2014, 15, 9, 3429-3439. [3] A.P. McCloskey, B.F. Gilmore and G.Laverty. Pathogens. 2014, 3, 791-821.

Fulfilling the 3Rs: Galleria mellonella as an In Vivo Model to Assess the Antimicrobial Efficacy of Self-assembling Peptide Hydrogels

The concept of ‘The Three Rs’ (The 3Rs: reduction, refinement and replacement) is an important consideration in the development of alternatives to animal testing in medical research. Invertebrate models such as Galleria mellonella are advantageous both economically and ethically.1 Galleria have proven to be effective alternatives to assess the antimicrobial activity of novel therapeutics.2
In this study Galleria mellonella are validated and used as an in vivo infection model to determine the antimicrobial activity of a novel self-assembling antimicrobial peptide NapFFKK.3 The peptide was considered as being non-toxic to the Galleria with 100% survival 120 hours post inoculation with NapFFKK. Following inoculation with Pseudomonas aeruginosa PAO1, Escherichia coli ATCC 11303, Staphylococcus epidermidis ATCC 35984 and Staphylococcus aureus ATCC 6538, the highest concentration allowing survival was selected and used as the test inoculum. Haemolymph was extracted from inoculated and peptide treated Galleria at either 24 or 72 hours post-treatment. Reduction in bacterial load was determined in comparison to a positive control. Bacterial load was decreased in all treated Galleria with decreasing antimicrobial activity demonstrated with a decreased concentration of peptide (2- log cycle reduction achieved in Escherichia coli inoculated Galleria treated with 2% NapFFKK). The results are promising regarding the use of Galleria mellonella as an infection model and NapFFKK as an effective novel antimicrobial.

Decoration of au and ag nanoparticles on self-assembling pseudopeptide-based nanofiber by using a short peptide as capping agent for metal nanoparticles

The surface of a nanofiber that is formed from a self-assembling pseudopeptide has been decorated by gold and silver nanoparticles that are stabilized by a dipeptide. Transmission electron microscopic images make the decoration visible. In this paper, a new strategy of mineralizing a pseudopeptide based nanofiber by gold and silver nanoparticles with use of a two-component nanografting method is described.

Formation of triple helical nanofibers using self-assembling chiral benzene-1,3,5-tricarboxamides and reversal of the nanostructure's handedness using mirror image building blocks

Intertwining triple helical nanofibers with an overall handedness have been formed from self-assembling chiral benzene-1,3,5-tricarboxamides 1, 2 and 3, whereas the achiralbenzene-1,3,5-tricarboxamide 4 upon self-association gives rise to straight nanofibers without any twist and transmission electron microscopy images of chiral compounds clearly demonstrate that the handedness of the triple helical nanofibers can be reversed by using the enantiomeric benzene-1,3,5-tricarboxamide building blocks.

The role of protecting groups in the formation of organogels through a nano-fibrillar network formed by self-assembling terminally protected tripeptides

A series of eight synthetic self-assembling terminally blocked tripeptides have been studied for gelation. Some of them form gels in various aromatic solvents including benzene, toluene, xylene, and chlorobenzene. It has been found that the protecting groups play an important role in the formation of organogels. It has been observed that, if the C-terminal has been changed from methyl ester to ethyl ester the gelation property does not change significantly (keeping the N-terminal protecting group same), while the change of the protecting group from ethyl ester to isopropyl ester completely abolishes the gelation property. Similarly, keeping the identical C-terminal protecting group (methyl ester) the results of the gelation study indicate that the substitution of N-terminal protection Boc-(tert-butyloxycarbonyl) to Cbz-(benzyloxycarbonyl) does change the gelation property insignificantly, while the change from Boc- to pivaloyl (Piv-) or acetyl (Ac-) group completely eliminates the gelation property. Morphological studies of the dried gels of two of the peptides indicate the presence of an entangled nano-fibrillar network that might be responsible for gelation. FTIR studies of the gels demonstrate that an intermolecular hydrogen bonding network is formed during gelation. Results of X-ray powder diffraction studies for these gelator peptides in different states (dried gels, gel, and bulk solids) reflected that the structure in the wet gel is distinctly different from the dried gel and solid state structures. Single crystal X-ray diffraction studies of a non-gelator peptide, which is structurally similar to the gelator molecules reveal that the peptide forms an antiparallel beta-sheet structure in crystals. (c) 2007 Elsevier Ltd. All rights reserved.

Low molecular weight organogelators from self-assembling synthetic tripeptides with coded amino acids: morphological, structural, thermodynamic and spectroscopic investigations

A series of self-assembling terminally blocked tripeptides (containing coded amino acids) form gels in various aromatic solvents including benzene, toluene, xylenes at low concentrations. However these tripeptides do not form gels in aliphatic hydrocarbons like n-hexane, cyclohexane, n-decane etc. Morphological studies of the dried gel indicate the presence of an entangled fibrous network, which is responsible for gelation. Differential scanning calorimetric (DSC) studies of the gels produced by peptide 1 clearly demonstrates thermoreversible nature of the gel and tripeptide-solvent complex may be produced during gel formation. FT-IR and H-1 NMR studies of the gels demonstrate that an intermolecular hydrogen-bonding network is formed during gelation. Single crystal X-ray diffraction studies for peptides 1, 2 and 3 have been performed to investigate the molecular arrangement that might be responsible for forming the fibrous network of these self-assembling peptide gelators. It has been found that the morph responsible for gelation of peptides 1, 2 and 3 in benzene is somewhat different from that of its xerogel.

Construction of Helical Nanofibers from Self-Assembling Pseudopeptide Building Blocks: Modulating the Handedness and Breaking the Helicity

Helical nanofibers are successfully constructed from suitable self-assembling pseudopeptide-based molecular building blocks. The handedness of these nanofibers can be reversed by using mirror-imaged pseudopeptide-based building blocks. Straight nanofibers are also constructed by modulating the molecular and supramolecular structures by the proper choice of the stereochemical nature of the molecular scaffolds. This study demonstrates that molecular structure and chirality are not the only determining factors for tuning the morphology and chirality of nanostructures; the nature of the supramolecular structures formed from the corresponding molecular scaffolds also plays a key role in dictating the shape and chirality of nanostructures. Helical nanofibers are suitable templates for fabricating dipeptide-capped gold nanoparticles, indicating a possible use of these nanofibers in the construction of arrays of gold nanoparticles.

Solvent-Assisted Formation of Vesicles by a Self-Assembling Ni-3-Schiff Base Complex

A linear trinuclear Ni-Schiff base complex [Ni-3(salpen)(2)(PhCH2COO)(2)(EtOH)] has been synthesized by combining Ni(ClO4)(2)center dot 6H(2)O, phenyl acetic acid (C6H5CH2COOH), and the Schiff base ligand, N,N'-bis(salicylidene)-1,3-pentanediamine (H(2)salpen). This complex is self-assembled through hydrogen bonding and C-H-g interaction in the solid state to generate a sheet-like architecture, while in organic solvent (CH2Cl2), it forms vesicles with a mean diameter of 290 nm and fused vesicles, depending upon the concentration of the solution. These vesicles act as an excellent carrier of dye molecules in CH2Cl2. The morphology of the complex has been determined by scanning electron microscopy and transmission electron microscopy experiments, and the encapsulation of dye has been examined by confocal microscopic image and electronic absorption spectra.

A short water-soluble self-assembling peptide forms amyloid-like fibrils

A water-soluble tripeptide Val-Ile-Ala (VIA) 1, bearing sequence identity with the C-terminal portion of the Alzheimer A beta-peptide (A beta(40-42)), self-assembles, in crystalline form, to produce an intermolecularly hydrogen bonded supramolecular beta-sheet structure which self-associates to form straight, unbranched nanofibrils exhibiting amyloid-like behavior; in contrast, the synthetic tripeptide Ala-Val-Ile (AVI) 2 self-assembles to produce a beta-sheet structure that forms branched nanofibrils which do not show any characteristic features of amyloid-like fibrils.