We report the design and synthesis of a biocompatible small-peptide-based compound for the controlled and targeted delivery of encapsulated bioactive metal ions through transformation of the internal nanostructures of its complexes. A tyrosine-based short-peptide amphiphile (sPA) was synthesized and observed to self-assemble into β-sheet-like secondary structures. The self-assembly of the designed sPA was modulated by application of different bioactive transition-metal ions, as was confirmed by spectroscopic and microscopic techniques. These bioactive metal-ion-conjugated sPA hybrid structures were further used to develop antibacterial materials. As a result of the excellent antibacterial activity of zinc ions the growth of clinically relevant bacteria such as Escherichia coli was inhibited in the presence of zinc⋅sPA conjugate. Bacterial testing demonstrated that, due to high biocompatibility with bacterial cells, the designed sPA acted as a metal ion delivery agent and might therefore show great potential in locally addressing bacterial infections.

Transition metal ions mediate the secondary structural transformation of hydrophobized sPA and can be applied to the design and development of stimuli-responsive nanomaterials.

The efficacy of protein-based medicines can be compromised by their rapid clearance from the blood circulatory system. Achieving optimal pharmacokinetics is a key requirement for the successful development of safe protein-based medicines. Protein PEGylation is a clinically proven strategy to increase the circulation half-life of protein-based medicines. One limitation of PEGylation is that there are few strategies that achieve site-specific conjugation of PEG to the protein. Here, we describe the covalent conjugation of PEG site-specifically to a polyhistidine tag (His-tag) on a protein. His-tag site-specific PEGylation was achieved with a domain antibody (dAb) that had a 6-histidine His-tag on the C-terminus (dAb-His(6)) and interferon α-2a (IFN) that had an 8-histidine His-tag on the N-terminus (His(8)-IFN). The site of PEGylation at the His-tag for both dAb-His(6)-PEG and PEG-His(8)-IFN was confirmed by digestion, chromatographic, and mass-spectral studies. A methionine was also inserted directly after the N-terminal His-tag in IFN to give His(8)Met-IFN. Cyanogen bromide digestion studies of PEG-His(8)Met-IFN were also consistent with PEGylation at the His-tag. By using increased stoichiometries of the PEGylation reagent, it was possible to conjugate two separate PEG molecules to the His-tag of both the dAb and IFN proteins. Stability studies followed by in vitro evaluation confirmed that these PEGylated proteins retained their biological activity. In vivo PK studies showed that all of the His-tag PEGylated samples displayed extended circulation half-lives. Together, our results indicate that site-specific, covalent PEG conjugation at a His-tag can be achieved and biological activity maintained with therapeutically relevant proteins.

We highlight the structural diversity of strategically designed two short peptide amphiphiles (sPAs) and describe their structure–function relationship studies. The shuffling of two key amino acids, that is, tyrosine and phenylalanine, in a designed sPA lead to a pair of constitutional isomers. Such small and strategic alteration can bring a substantial change in the self‐assembling pattern. Inspired from the naturally occurring metallopeptides, bioactive transition‐metal ions were used for constructing the unusual nanostructures. Use of appropriate metal ions created bigger differences between the properties of these isomers and hence the self‐assembly. Coordination of appropriate transition metal ions modifies the internal nanoscale structures of sPA, thus leading to the formation of vertically stacked terraced layers with decreasing size, which possess a high degree of dimensional regularity. We propose that such metal‐induced terraced nanodome‐like hierarchical self‐assembly may have relevance for specific biotechnology applications.

We report the aggregation and photophysical properties of a pyridothiazole-based, aggregation-induced, emission-enhancement (AIEE) luminogen 4-(5-methoxy-thiazolo­[4,5-b]­pyridin-2-yl)­benzoic acid (PTC1) and its application for the sensitive detection and monitoring of amyloid fibrillation. The aggregation properties of the AIEE probe were extensively studied by atomic force microscopy (AFM) and dynamic light scattering (DLS), and it was noted that as aggregation increases the fluorescence of PTC1 also was increased. The fluorescence of PTC1 was quenched upon the addition of cupric (Cu2+) ions, while the fluorescence is regenerated in the presence of amyloid fibers. AFM studies reveal that the PTC1 molecules self-associate/aggregate to hairy micelle-like structures, which dissociate or disrupt in the presence of the Cu2+ ions and again reassemble in the presence of amyloid fibers. Hence, the quenching and regeneration of PTC1 fluorescence may be attributed to the disaggregation and aggregation-induced emission (AIE), respectively. Further, a comparative analysis of the performance of PTC1 was done with conventional Thioflavin T, which confirms it to be a more sensitive probe for the detection of the amyloid, both in the presence and absence of Cu2+ ions. The experimental results were also validated theoretically via molecular docking and simulation studies. Of note, a very simple, facile, and cost-effective methodology for the detection of the amyloid fibers is presented, wherein fluorescence quenching/enhancement can be visualized under the UV light without the use of sophisticated instrumentation techniques. The AIEE probe was designed using an unusual pyridothiazole scaffold unlike commonly used archetypal AIE scaffolds based on tetraphenylethene (TPE) and hexaphenylsilole (HPS). Hence, the work also has implications in designing future AIEE dyes based on the pyridothiazole scaffold reported.