Self-assembly Of Oligopeptides Research Articles

Weak Alignment Liquid Crystal Media from Colloidal Dispersion and Self-assembled Oligopeptide for Anisotropic NMR

Weak Alignment Liquid Crystal Media from Colloidal Dispersion and Self-assembled Oligopeptide for Anisotropic NMR

Comparative Efficacy of Novel Biomimetic Remineralising Technologies.

Biomimetic technologies for the remineralisation of enamel subsurface lesions (ESLs) have been developed and include: fluorocalcium phosphosilicate bioglass (BG/F); casein phosphopeptide-amorphous calcium phosphate (CPP−ACP) and with fluoride (CPP−ACFP); and self-assembling oligopeptide P11-4 (SAP). The aim of this study was to compare the remineralisation of ESLs in vitro using these technologies. Human enamel slabs with ESLs were cut into two half-slabs; one half-slab was untreated (control), and the other half was treated by exposure to one of the four technologies with artificial saliva (AS) or AS alone for 14 days at 37 °C. The technologies were applied to the ESL surface according to the manufacturer’s instructions. At the completion of each treatment, the treated half-slabs and their paired control half-slabs were embedded, sectioned and the mineral content was determined using transverse microradiography. The change in mineral content (remineralisation) between treatments was statistically analysed using one-way ANOVA. The order from highest to lowest remineralisation was CPP−ACFP (52.6 ± 2.6%) > CPP−ACP (43.0 ± 4.9%) > BG/F (13.2 ± 2.5%) > SAP (5.8 ± 1.6%) > AS (2.1 ± 0.5%). Only CPP−ACFP and CPP−ACP produced remineralisation throughout the body of the lesions. All four biomimetic technologies had some effect on the remineralisation of ESLs; however, CPP−ACFP with calcium, phosphate and fluoride ions stabilised by CPP was superior in the level and pattern of remineralisation obtained.

Self-Assembled Oligopeptide (FK)4 as a Chiral Alignment Medium for the Anisotropic NMR Analysis of Organic Compounds.

Anisotropic NMR parameters have been proven to be powerful for the structural elucidation of organic molecules. Herein, we present an alignment medium based on the self-assembled (FK)4 oligopeptide, showing excellent properties in measurements of anisotropic NMR parameters in both D2O and CD3OD. The preparation of the (FK)4-based alignment medium is simple and rapid. The low viscosity of the anisotropic phase makes it easy to be transferred to the NMR tube. The alignment of the oligopeptide is fast, stable, and homogeneous, with weak background signals, permitting the acquirement of high-quality NMR spectra. The performance of this alignment medium in residual dipolar coupling measurements and diastereomer discriminations is demonstrated by analyzing several different analytes. The enantiodiscrimination property of the (FK)4 oligopeptide is revealed by the difference of residual chemical shift anisotropy of the two enantiomers in the 1D 13C spectrum, granting its potential use for the quantification and identification of enantiomers of small molecules.

Evaluation of Children Caries Risk Factors: A Narrative Review of Nutritional Aspects, Oral Hygiene Habits, and Bacterial Alterations.

Dental caries is one of the most common diseases—both in adults and children—that occurs due to the demineralization of enamel and dentine by the organic acids formed from bacteria present in dental plaques through anaerobic metabolism of dietary sugars. The aim of this article is to provide a guideline to assess the risk of caries by looking for the main factors involved. Literature research was performed for studies that analyzed the factors most involved in the development of child caries, such as poor oral hygiene, bad eating habits (or food disorders), and an alteration of the oral bacterial flora—with an increase of Streptococci spp., Lactobacilli spp., Candida albicans, Cryptococcus neoformans, and Candida sake. It is therefore essential to assess the risk of caries in children, based on the assessment of risk factors, in order to be able to establish preventive and/or therapeutic approaches that will reduce or stop the development of dental caries. The use of fluoride products, products made from casein phosphopeptide-amorphous calcium phosphate, substituted zinc biomimetic hydroxyapatite products, or products containing self-assembling oligopeptide SAP-P11-4 are useful. In terms of the clinical approach, a communicative approach should be added to learn about the eating habits and the oral hygiene habits of the child and parents; in addition, the use of a simple method to frame the factors involved, and subsequently establish the risk of carious lesions in the child, allows the reduction of the DMFT (Decayed Missing Filled Teeth) or ICDAS (International Caries Detection and Assessment System) index on large scales.

Programmable alignment media from self-assembled oligopeptide amphiphiles for the measurement of independent sets of residual dipolar couplings in organic solvents.

NMR spectroscopy in anisotropic media has emerged as a powerful technique for the structural elucidation of organic molecules. Its application requires weak alignment of analytes by means of suitable alignment media. Although a number of alignment media, that are compatible with organic solvents, have been introduced in the last 20 years, acquiring a number of independent, non-linearly related sets of anisotropic NMR data from the same organic solvent system remains a formidable challenge, which is however crucial for the alignment simulations and deriving dynamic and structural information of organic molecules unambiguously. Herein, we introduce a programmable strategy to construct several distinct peptide-based alignment media by adjusting the amino acid sequence, which allows us to measure independent sets of residual dipolar couplings (RDCs) in a highly efficient and accurate manner. This study opens a new avenue for de novo structure determination of organic compounds without requiring prior structural information.

Spider silk-inspired peptide multiblock hybrid copolymers for self-healable thin film materials

Spider silk-inspired peptide multiblock hybrid copolymers composed of alternately aligned self-assembling oligopeptides and flexible polypropylene glycol were newly designed for functional nano/microfilm materials.

Enzyme-instructed hybrid nanogel/nanofiber oligopeptide hydrogel for localized protein delivery

Enzyme-catalysis self-assembled oligopeptide hydrogel holds great interest in drug delivery, which has merits of biocompatibility, biodegradability and mild gelation conditions. However, its application for protein delivery is greatly limited by inevitable degradation of enzyme on the encapsulated proteins leading to loss of protein activity. Moreover, for the intracellularly acted proteins, cell membrane as a primary barrier hinders the transmembrane delivery of proteins. The internalized proteins also suffer from acidic and enzymatic degradation in endosomes and lysosomes. We herein develop a protease-manipulated hybrid nanogel/nanofiber hydrogel for localized delivery of intracellularly acted proteins. The embedded polymeric nanogels (CytoC/aNGs) preserve activity of cytochrome c (CytoC) that is an intracellular activator for cell apoptosis as a model protein against proteolysis, and do not affect the gelation properties of the protease-catalysis assembled hydrogels. The injectable hydrogel (CytoC/aNGs/Gel) serves as a reservoir to enhance intratumoral retention and realize sustainable release of CytoC/aNGs. The released CytoC/aNGs increase cellular uptake of CytoC and enhance its intracellular delivery to its target site, cytoplasm, resulting in favorable apoptosis-inducing and cytotoxic effects. We show that a single local administration of CytoC/aNGs/Gel efficiently inhibit the tumor growth in the breast tumor mouse model.

A novel amphiphilic oligopeptide induced the intrafibrillar mineralisation via interacting with collagen and minerals.

Mineralised collagen fibrils constitute the basic building blocks of bone, dentin and cementum. Noncollagenous proteins (NCPs) that are indispensable for collagen biomineralisation are not commercially available, and the mechanism of intrafibrillar mineralisation remains debatable. Herein, synthetic biomimetic molecules are regarded as alternative candidates for NCPs, and more convenient in revealing the mechanism of intrafibrillar mineralisation in vitro. Here, we fabricated a novel amphiphilic oligopeptide imitating a natural NCP. We aimed to investigate the effectiveness of the oligopeptide in intrafibrillar mineralisation and partially reveal the corresponding mechanism in vitro. The effectiveness of the oligopeptide in intrafibrillar mineralisation was characterised from the following aspects: (1) mineral interaction, (2) collagen binding and (3) induction of intrafibrillar mineralisation. Results indicated that the self-assembled oligopeptide could attract calcium ions inducing the formation of amorphous precursors; and bind onto the surface of collagen fibrils. These processes were mainly driven by the electrostatic attraction and hydrogen bonds. The self-assembled oligopeptide induced the intrafibrillar mineralisation of reconstituted collagen fibrils, in which the c-axis of apatite crystallites was roughly parallel to the long axis of the fibrils. The collagen mineralisation was achieved by binding with the self-assembled oligopeptide to increase the pool of mineralization precursors available for intrafibrillar mineralisation. In addition, the self-assembled oligopeptide induced dentin collagen remineralisation and formed a 30 μm-thick remineralised layer within 96 h. Our work sheds light on the fabrication of a novel biomimetic molecule for collagen mineralisation. The results should serve as a reference for understanding the mechanism of intrafibrillar mineralisation.

One-step maltose-functionalization of magnetic nanoparticles based on self-assembled oligopeptides for selective enrichment of glycopeptides

Magnetic nanoparticles (MNPs) have been widely explored in enrichment of low-abundance glycoproteins/glycopeptides prior to mass spectrometry analysis in glycoproteomics. Currently, most functional groups for recognizing glycoproteins/glycopeptides are usually immobilized on the nanomaterial surface based on covalent modification, which suffers from multistep treatment, surface-dependence, and harsh conditions. In this work, we first report a facile and rapid method for surface functionalization and subsequent glycopeptides enrichment via one-step assembly of maltose-modified oligopeptides with a sequence of Ala-Glu-Ala-Glu-Ala-Lys-Ala-Lys (AEK8-maltose). In physiological conditions, AEK8-maltose readily self-organized into a complete coating layer dominated by β-sheets on the surface of SiO2@Fe3O4 and C@Fe3O4 MNPs, which remain intact to repeat washing with acidic organic and aqueous solutions extensively used in the sample enrichment treatments. Thus, the resulting AEK8-maltose functionalized MNPs show excellent performance in enrichment of glycopeptides in standard glycoprotein digests (24 glycopeptides from horseradish peroxidase (HRP), 31 glycopeptides from immunoglobulin (IgG)) and human serum digests (282 glycopeptides), including rapid enrichment speed (5 min), high detection sensitivity (0.001 ng/μL HRP), high selectivity (mass ratios of HRP and bovine serum albumin (BSA) digests up to 1:150), good enrichment recovery (over 86.3%), remarkable stability (repeatable for more than 8 times), and excellent renewability, which are better than or comparable with the literature results reported to date. The current work based on self-assembling oligopeptides provides a mild, economic and nontoxic procedure for one-step surface functionalization of various nanomaterials.

Evidence for Prenucleated Fibrilogenesis of Acid-Mediated Self-Assembling Oligopeptides via Molecular Simulation and Fluorescence Correlation Spectroscopy.

An important step in controlling biomimetic amyloid systems is understanding the self-assembly reaction kinetics. We are interested in a family of such materials characterized by symmetric sequences of amino acids flanking a π-conjugated functional core. Many of these materials rapidly self-assemble into long fibers upon protonation in an acidic environment. Despite extensive investigation of these materials' properties, little is yet understood regarding their reaction kinetics. Based on previous studies, we have chosen DFAG-4T-GAFD as a representative system and conducted molecular dynamics simulations to show that although large-scale assembly is induced by lowering pH, some degree of assembly is thermodynamically favorable in high-pH nonprotonating environments. These results are consistent with findings for other systems such as DFAG-OPV-GAFD. The nonprotonated aggregation also appears to be concentration dependent, occurring at concentrations of 100 nM and above. Single molecule measurements using fluorescence correlation spectroscopy provide experimental support for these computational predictions. We find evidence of spontaneous aggregation in aqueous solutions of peptides with concentrations as low as 100 nM; however, 10 nM solutions appear to be largely homogeneous solutions of unassembled monomer. These results indicate that the simplest explanations for kinetics of acid-mediated assembly-protonation-induced nucleation by monomeric addition followed by subsequent stages of aggregation and elongation-are inappropriate in this system. In fact, the system only exists as pure monomer in very low concentrations, nucleation actually occurs in the absence of protonating elements at concentrations typically used for experiments, and pH triggered assembly proceeds from these preassembled aggregates. Accordingly, triggered assembly must be considered to operate outside the domain of nucleation-dependent models.

Machine learning and molecular design of self-assembling -conjugated oligopeptides

Self-assembling oligopeptides present a means to fabricate biocompatible supramolecular aggregates with engineered electronic and optical functionality. We conducted molecular dynamics simulations of self-assembling synthetic oligopeptides with Asp-X-X-X--X-X-X-Asp architectures. Dimerisation and trimerisation free energies were computed for a range of Asp-X-X-X amino acid sequences, and for perylenediimide (PDI) and naphthalenediimide (NDI) conjugated cores that mediate hydrophobic stacking and electron delocalisation within the self-assembled nanostructure. The larger PDI cores elevated oligomerisation free energies by a factor of 2-3 relative to NDI and also improved alignment of the oligopeptides within the stack. Training of a quantitative structure–property relationship (QSPR) model revealed key physicochemical determinants of the oligomerisation free energies and produced a predictive model for the oligomerisation thermodynamics. Oligopeptides with moderate dimerisation and trimerisation free energies of (-25) produced aggregates with the best in-register parallel stacking, and we used this criterion within our QSPR model to perform high-throughput virtual screening to identify promising candidates for the spontaneous assembly of ordered nanoaggregates. We identified a small number of oligopeptide candidates for direct testing in large scale molecular simulations, and predict a novel chemistry DAVG-PDI-GVAD previously unstudied by experiment or simulation to produce well-aligned nanoaggregates expected to possess good optical and electronic functionality.

Biofunctionalization of TiO2 surfaces with self-assembling oligopeptides in different pH and Ionic Strength conditions: Charge effects and molecular organization

Self-assembling peptides (SAPs) were investigated by means of XPS and Angular Dependent NEXAFS spectroscopies, with the aim to probe the influence of pH and Ionic Strength conditions on the chemical structure and molecular organization of SAPs anchored on titania surfaces. XPS at the C1s, N1s, O1s core levels allowed to study surfaces and biomolecule/substrate interfaces. NEXAFS data allowed ascertaining that SAPs molecular structure is preserved upon grafting to the titania surface. Angular Dependent NEXAFS was used to investigate the influence of environmental conditions on the molecular organization behaviour. The objective of our study was to establish a set of methodologies for obtaining arrangements of well-organized biomolecules on scaffolds surfaces as a basic technology to develop and optimize cells adhesion and proliferation for tissue engineering applications.

A Self‐Assembled Oligopeptide as a Versatile NMR Alignment Medium for the Measurement of Residual Dipolar Couplings in Methanol

AbstractResidual dipolar coupling (RDC) is a powerful structural parameter for the determination of the constitution, conformation, and configuration of organic molecules. Herein, we report the first liquid crystal‐based orienting medium that is compatible with MeOH, thus enabling RDC acquisitions of a wide range of intermediate to polar organic molecules. The liquid crystals were produced from self‐assembled oligopeptide nanotubes (AAKLVFF), which are stable at very low concentrations. The presented alignment medium is highly homogeneous, and the size of RDCs can be scaled with the concentration of the peptide. To assess the accuracy of the RDC measurement by employing this new medium, seven bioactive natural products from different classes were chosen and analyzed. The straightforward preparation of the anisotropic alignment sample will offer a versatile and robust protocol for the routine RDC measurement of natural products.

A Self-Assembled Oligopeptide as a Versatile NMR Alignment Medium for the Measurement of Residual Dipolar Couplings in Methanol.

Residual dipolar coupling (RDC) is a powerful structural parameter for the determination of the constitution, conformation, and configuration of organic molecules. Herein, we report the first liquid crystal-based orienting medium that is compatible with MeOH, thus enabling RDC acquisitions of a wide range of intermediate to polar organic molecules. The liquid crystals were produced from self-assembled oligopeptide nanotubes (AAKLVFF), which are stable at very low concentrations. The presented alignment medium is highly homogeneous, and the size of RDCs can be scaled with the concentration of the peptide. To assess the accuracy of the RDC measurement by employing this new medium, seven bioactive natural products from different classes were chosen and analyzed. The straightforward preparation of the anisotropic alignment sample will offer a versatile and robust protocol for the routine RDC measurement of natural products.

寡肽自组装纳米材料研究进展

Due to the mellow synthetic methods, determined molecular structures, well biocompatibility and controlled degradation, oligopeptides as the self-assembled units have attracted increasing research interest, and their self-assemblies present a great potential in biomedical fields such as drug/gene delivery, 3D cell culture, tissue engineering scaffold and biomineralization. This review summarized the research progress in self-assembly of oligopeptides including linear and cyclic peptides in recent years. The molecular design, self-assembled morpho- logies, proposed mechanisms, as well as their biomedical applications are reviewed in detail.

Variation of formal hydrogen-bonding networks within electronically delocalized π-conjugated oligopeptide nanostructures.

This photophysical study characterizes the generality of intermolecular electronic interactions present within nanomaterials derived from self-assembling oligopeptides with embedded π-conjugated oligophenylenevinylene (OPV) subunits stilbene and distyrylbenzene that in principle present two distinct β-sheet motifs. Two different synthetic approaches led to oligopeptides that upon self-assembly are expected to self-assemble into multimeric aggregates stabilized by β-sheet-like secondary structures. The target molecules express either two C-termini linked to the central OPV core (symmetric peptides) or the more common N-termini to C-termini polarity typical of natural oligopeptides (nonsymmetric peptides). Both peptide secondary structures were shown to form extended 1-D peptide aggregates with intimate intermolecular π-electron interactions. Differences in length of the π-conjugated OPV segments resulted in differing extents of intermolecular interactions and the resulting photophysics. The peptides containing the shorter stilbene (OPV2) units showed little ground state interactions and resulted in excimeric emission, while the longer distyrylbenzene (OPV3) peptides had different ground state interactions between adjacent π-conjugated subunits resulting in either perturbed electronic properties arising from exciton coupling or excimer-like excited states. Molecular dynamics simulations of nascent aggregate formation predict peptide dimerization to be a spontaneous process, possessing thermodynamic driving potentials in the range 2-6 kcal/mol for the four molecules considered. Antiparallel stacking of the peptides containing an OPV3 subunit is thermodynamically favored over the parallel orientation, whereas both arrangements are equally favored for the peptides containing an OPV2 subunit. This study validates the generality of peptide-π-peptide self-assembly to provide electronically delocalized supramolecular structures and suggests flexibility in peptide sequence design as a way to tune the material properties of π-conjugated supramolecular polymers.

A novel self-assembled oligopeptide amphiphile for biomimetic mineralization of enamel.

BackgroundResearchers are looking for biomimetic mineralization of ena/mel to manage dental erosion. This study evaluated biomimetic mineralization of demineralized enamel induced by a synthetic and self-assembled oligopeptide amphiphile (OPA).ResultsThe results showed that the OPA self-assembled into nano-fibres in the presence of calcium ions and in neutral acidity. The OPA was alternately immersed in calcium chloride and sodium hypophosphate solutions to evaluate its property of mineralization. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) showed nucleation and growth of amorphous calcium phosphate along the self-assembled OPA nano-fibres when it was repetitively exposed to solutions with calcium and phosphate ions. Energy dispersive spectrometry (EDS) confirmed that these nano-particles contained calcium and phosphate. Furthermore, electron diffraction pattern suggested that the nano-particles precipitated on OPA nano-fibres were comparable to amorphous calcium phosphate. Acid-etched human enamel slices were incubated at 37°C in metastable calcium phosphate solution with the OPA for biomimetic mineralization. SEM and X-ray diffraction indicated that the OPA induced the formation of hydroxyapatite crystals in organized bundles on etched enamel. TEM micrographs revealed there were 20–30 nm nano-amorphous calcium phosphate precipitates in the biomimetic mineralizing solution. The particles were found separately bound to the oligopeptide fibres. Biomimetic mineralization with or without the oligopeptide increased demineralized enamel microhardness.ConclusionsA novel OPA was successfully fabricated, which fostered the biomimetic mineralization of demineralized enamel. It is one of the primary steps towards the design and construction of novel biomaterial for future clinical therapy of dental erosion.

Self-assembled peptide nanoparticles as tumor microenvironment activatable probes for tumor targeting and imaging

Design of specific and sensitive imaging probes for targeting tumor microenvironment holds great promise to achieve precise detection and rapid responsiveness to neoplastic tissues. Dysregulated pH, one of the most remarkable hallmarks of tumor microenvironment, can be considered as a good specific trigger for the design of broad-spectrum and local-environment responsive imaging probes. However, the current existing design strategies for pH-responsive systems are insufficient to meet the needs for a rapid and tumor-specific diagnosis. Here we reported a novel biomimetic nanostructure based on oligopeptide self-assembly that can quickly switch into dissociated stage with active fluorescence property from self-assembled stage with quenched fluorescence activity when encountering a subtle pH-change in tumor microenvironment (pH6.8 vs. 7.4). This oligopeptide-assembly is examined as tumor microenvironment activatable probes for both intratumoral and intravenous in vivo tumor imaging. Through the distinct fluorescent intensities, it is validated that the acidic tumor microenvironment can activate stronger fluorescence signals. The tailor-made self-assembled oligopeptide nanomaterials have the potential for efficient and specific in situ diagnosis of various solid tumors with a weakly acidic microenvironment, which is expected to be of crucial importance for clinical tumor diagnostics.

Mechanical characterization of self‐assembling peptide hydrogels by microindentation

Hydrogels formed from self-assembling synthetic oligopeptides have been studied for almost 2 decades for use in tissue engineering and drug delivery. Although a great deal has been learned about the microstructure of these materials, there remain questions about how peptide filaments are ordered to form a gel. These unanswered questions leave a disconnect between our understanding of the observed nanoscale mechanical properties of peptide filaments and the macroscale properties of the gels they constitute. This study helps to bridge this gap by examining the role of filament length and interfilament crosslinks in determining bulk mechanical properties. Microindentation was used for mechanical characterization, and microstructure was observed using thin-section transmission electron microscopy images of gels embedded in resin. Results suggest a gel structure in which filaments are not densely bundled as previously suggested and confirm that crosslinking can be an effective strategy to increase the stiffness of self-assembling oligopeptide gels.

Anomalous Conformational Instability and Hydrogel Formation of a Cationic Class of Self-Assembling Oligopeptides

A detailed understanding of the mechanistic principles which govern peptide and protein self-assembly is of considerable biomedical and biotechnological importance. Owing to the diversity of peptide and protein sequences which have been shown to aggregate into ordered structures, the ability to self-assemble is now recognized as an inherent feature of the polypeptide backbone. It is therefore of utmost importance to elucidate the rules governing the self-assembly process. De novo designed oligopeptides, based on sequences of alternating hydrophilic and hydrophobic residues and containing complementary charge distributions, have shown the potential to self-assemble into hydrogels rich in β-sheet secondary structure. Here we present and characterize the unexpected self-assembly and hydrogel formation of AK-16, an alanine-rich oligopeptide, whose sequence does not abide by typical rules which allow for peptide self-organization. AK-16 spontaneously forms soluble, thermodynamically unstable β-sheet-rich aggregates, which can be stabilized by salt addition to yield a self-supporting macroscopic hydrogel. The AK-16 hydrogel exhibits the ability to encapsulate and slowly release a model protein. This one-charge-type system represents a novel class of self-assembling oligopeptides, in which the initial conformational instability can be exploited to tune the viscosity and physicochemical properties of the resultant hydrogel. This study provides insight for the future de novo design of self-assembling oligopeptides.