Alternative Assembly of α-Synuclein Leading to Protein Film Formation and Its Application for Developing Polydiacetylene-Based Sensing Materials.

Understanding the self-assembly process of amyloidogenic protein is valuable not only to find its pathological implication but also to prepare protein-based biomaterials. α-Synuclein (αS), a pathological component of Parkinson's disease, producing one-dimensional (1D) amyloid fibrils, has been employed to generate two-dimensional (2D) protein films by encouraging an alternative self-assembly process. At a high temperature of 50 °C, αS molecules self-assembled into 2D films instead of 1D amyloid fibrils, whereas the fibrils were the major product at 37 °C. Based on circular dichroism and Fourier transform infrared spectroscopy analyses, the film was produced via a structural transition from the initial random to still undefined but mostly the turn or loop structure, which was distinctive from the ß-sheet formation observed with the amyloid fibrils. The αS 2D film was also routinely prepared at the oil-water interface and used as a matrix to produce polydiacetylene-based sensing materials. 10,12-Pentacosadiynoic acids (PCDA) were aligned on the film and photopolymerized to form a π-conjugated molecular assembly yielding a blue color. Its colorimetric transition to red was induced by increasing the temperature. This functionalized protein film increased its height from 40 to 55 nm upon PCDA immobilization and exhibited enhanced physical and chemical stability. In addition, the modified film showed remarkably high electrical conductivity only in the red state. This film, therefore, can be considered as a robust protein-based hybrid biomaterial capable of simultaneously recognizing various external stimuli (heat, pH, and solvents) with changes in color and conductivity, and it is expected to be utilized as a basic material for the development of biocompatible sensors.

Fabrication of Protease-Sensitive and Light-Responsive Microcapsules Encompassed with Single Layer of Gold Nanoparticles by Using Self-Assembly Protein of α-Synuclein.

A bioapplicable cargo delivery system requires the following characteristics of biocompatibility, in vivo stability, and selective cargo release at target sites. We introduce herein the microcapsules enclosed with a single-layered shell of gold nanoparticles (AuNPs) mutually connected by an amyloidogenic protein of α-synuclein (αS). The microcapsules were fabricated by producing oil(chloroform)-in-water Pickering emulsions of the αS-encapsulated AuNPs and subsequent molecular engagement of the outlying αS molecules, leading to formidable ß-sheet formation in the presence of chloroform. The wrinkled skin of microcapsules obtained after evaporation of the internal chloroform also reflects robustness of the protein-protein interaction, which was experimentally confirmed by their rheological stability. For the emulsions loaded with rhodamine 6G, their dye release was demonstrated to be controlled by proteases. Along with their photothermal activity, the AuNP-containing microcapsules and their proteolyzed fragments were therefore suggested to be capable of eliminating aberrant cells in the protease-activated pathologically affected areas. Orthogonal cargo loading was also achieved by encapsulating both hydrophobic and hydrophilic substances either directly dissolved in chloroform or prepackaged in inverted micelles, respectively. Microcapsule's functionality was further expanded by localizing quantum dots, magnetic nanoparticles, and antibodies inside or on the surface of the microcapsules. Taken together, these multimodal AuNP microcapsules are suggested to be an ideal cargo carrier system, which could be employed in not only biomedical theranostic applications as they exhibit structural robustness, specific targeting, triggered release, and photothermal activity but also sensor development in general.

Exposure to bacterial endotoxin generates a distinct strain of α-synuclein fibril.

A single amyloidogenic protein is implicated in multiple neurological diseases and capable of generating a number of aggregate "strains" with distinct structures. Among the amyloidogenic proteins, α-synuclein generates multiple patterns of proteinopathies in a group of diseases, such as Parkinson disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). However, the link between specific conformations and distinct pathologies, the key concept of the strain hypothesis, remains elusive. Here we show that in the presence of bacterial endotoxin, lipopolysaccharide (LPS), α-synuclein generated a self-renewable, structurally distinct fibril strain that consistently induced specific patterns of synucleinopathies in mice. These results suggest that amyloid fibrils with self-renewable structures cause distinct types of proteinopathies despite the identical primary structure and that exposure to exogenous pathogens may contribute to the diversity of synucleinopathies.

Radiating amyloid fibril formation on the surface of lipid membranes through unit-assembly of oligomeric species of α-synuclein.

BACKGROUND: Lewy body in the substantia nigra is a cardinal pathological feature of Parkinson's disease. Despite enormous efforts, the cause-and-effect relationship between Lewy body formation and the disorder is yet to be explicitly unveiled. METHODOLOGY/PRINCIPAL FINDINGS: Here, we showed that radiating amyloid fibrils (RAFs) were instantly developed on the surface of synthetic lipid membranes from the ß-sheet free oligomeric species of α-synuclein through a unit-assembly process. The burgeoning RAFs were successfully matured by feeding them with additional oligomers, which led to concomitant dramatic shrinkage and disintegration of the membranes by pulling off lipid molecules to the extending fibrils. Mitochondria and lysosomes were demonstrated to be disrupted by the oligomeric α-synuclein via membrane-dependent fibril formation. CONCLUSION: The physical structure formation of amyloid fibrils, therefore, could be considered as detrimental to the cells by affecting membrane integrity of the intracellular organelles, which might be a molecular cause for the neuronal degeneration observed in Parkinson's disease.