Secondary Structure Properties Research Articles

A Coronin 1-Derived Peptide Inhibits Membrane Fusion by Modulating Membrane Organization and Dynamics.

Membrane fusion is considered the first step in the entry of enveloped viruses into the host cell. Several targeted strategies have been implemented to block viral entry by limiting the fusion protein to form a six-helix bundle, which is a prerequisite for fusion. Nonetheless, the development of broad-spectrum fusion inhibitors is essential to combat emerging and re-emerging viral infections. TG-23, a coronin 1, a tryptophan-aspartate-rich phagosomal protein-derived peptide, demonstrated inhibition of fusion between small unilamellar vesicles (SUVs) by modulating the membrane's physical properties. However, its inhibitory efficacy reduces with an increasing concentration of membrane cholesterol. The present work aims to develop a fusion inhibitor whose efficacy would be unaltered in the presence of membrane cholesterol. A stretch of the tryptophan-aspartic acid-containing peptide with a similar secondary structure and hydrophobicity profile of TG-23 from coronin 1 was synthesized, and its ability to inhibit SUV-SUV fusion with varying concentrations of membrane cholesterol was evaluated. Our results demonstrate that the GG-21 peptide inhibits fusion irrespective of the cholesterol content of the membrane. We have further evaluated the peptide-induced change in the membrane organization and dynamics utilizing arrays of steady-state and time-resolved fluorescence measurements and correlated these results with their effect on fusion. Interestingly, GG-21 displays inhibitory efficacy in a wide variety of lipid compositions despite having a secondary structure and physical properties similar to those of TG-23. Overall, our results advocate that the secondary structure and physical properties of the peptide may not be sufficient to predict its inhibitory efficacy.

Multiscale Simulations of Self-Assembling Peptides: Surface and Core Hydrophobicity Determine Fibril Stability and Amyloid Aggregation.

Assemblies of peptides and proteins through specific intermolecular interactions set the basis for macroscopic materials found in nature. Peptides provide easily tunable hydrogen-bonding interactions, which can lead to the formation of ordered structures such as highly stable β-sheets that can form amyloid-like supramolecular peptide nanofibrils (PNFs). PNFs are of special interest, as they could be considered as mimics of various fibrillar structures found in nature. In their ability to serve as supramolecular scaffolds, they could mimic certain features of the extracellular matrix to provide stability, interact with pathogens such as virions, and transduce signals between the outside and inside of cells. Many PNFs have been reported that reveal rich bioactivities. PNFs supporting neuronal cell growth or lentiviral gene transduction have been studied systematically, and their material properties were correlated to bioactivities. However, the impact of the structure of PNFs, their dynamics, and stabilities on their unique functions is still elusive. Herein, we provide a microscopic view of the self-assembled PNFs to unravel how the amino acid sequence of self-assembling peptides affects their secondary structure and dynamic properties of the peptides within supramolecular fibrils. Based on sequence truncation, amino acid substitution, and sequence reordering, we demonstrate that peptide-peptide aggregation propensity is critical to form bioactive β-sheet-rich structures. In contrast to previous studies, a very high peptide aggregation propensity reduces bioactivity due to intermolecular misalignment and instabilities that emerge when fibrils are in close proximity to other fibrils in solution. Our multiscale simulation approach correlates changes in biological activity back to single amino acid modifications. Understanding these relationships could lead to future material discoveries where the molecular sequence predictably determines the macroscopic properties and biological activity. In addition, our studies may provide new insights into naturally occurring amyloid fibrils in neurodegenerative diseases.

Gelation Behavior and Drug Sustained-Release Properties of a Helix Peptide Organohydrogel with pH Responsiveness.

Based on the typical similar repeat units (abcdefg)n of α-helical structure, the peptide H was designed to self-assemble into an organohydrogel in response to pH. Depending on the different pH, the proportions of secondary structure, microstructure, and mechanical properties of the gel were investigated. Circular dichroism (CD) and Fourier transform infrared (FT-IR) showed that the proportion of α-helical structure gradually increased to become dominant with the increase of pH. Combining transmission electron microscopy (TEM) and atomic force microscopy (AFM), it was found that the increase of the ordered α-helix structure promoted fiber formation. The further increase in pH changed the intermolecular forces, resulting in an increase in the α-helix content and the enhancement of helix-helix interaction, causing the gel fibers to converge into thicker and more dense ones. The temperature test showed the stable rheological properties of the organohydrogel between 20-60 °C. Drug release and cytotoxicity showed that the DOX-loaded organohydrogel could have a better release in an acidic environment, indicating its potential application as a drug local delivery carrier.

Incorporation of nanoparticles in silk fibroin solution: Toward water‐stable silk fibroin films possessing silk I structure

AbstractWater‐stable silk fibroin (SF) films are normally prepared by transforming the secondary structure of SF, from amorphous silk I (random coils) to silk II (β‐sheet), via physical or chemical means. Although mechanically strong, these films are generally brittle. Herein, we prepared water‐stable flexible SF films possessing silk I rather than silk II structure. Films were prepared by casting SF solution possessing SF nanoparticles induced by two different methods: (1) pre‐prepared SF nanoparticles by ethanol were added into SF solution; (2) nanoparticles were induced in SF solution via autoclaving. These films were compared with each other and with water‐annealed and methonol‐annealed films for their secondary structure and physical properties. The as‐prepared films were primarily composed of silk I structure, as validated by x‐ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy. However, the water‐annealed and methanol‐annealed films exhibited silk II structure. The films prepared by induction of nanoparticles were water‐stable, transparent, mechanically strong (tensile strength as high as 5.14 ± 1.43 MPa), exhibited tunable degradation and sustained drug release properties. As for mechanical properties, these films displayed 4 times and 12 times improved mechanical ductility as compared with water‐annealed and methanol‐annealed films, respectively. Moreover, these films exhibited fast enzymatic degradation as compared with their counterparts with silk II structure. These films hold great promise as a biomaterial for tissue engineering applications especially where flexibility and fast degradation of scaffold are important.

Characterization of O/W emulgels based on whey protein-alginate-inulin coacervates: Influence of temperature and ultrasound as protein preconditioning process

Preconditioning processes in proteins play a crucial role in enhancing their functional properties as surface active agents. Whey protein isolate (WPI, 20 wt%) was preconditioned via temperature (WPIT, 90 °C) or ultrasound (WPIUS, 20 kHz, 80 % amplitude). FTIR and zeta potential analysis demonstrated the effect of the preconditioning process on the secondary structure and surface properties of WPI. WPI-Alginate:Inulin (AI) complex coacervates (CCWPI:AI) were formed at pH 3.0 using WPIT and WPIUS, and the associative electrostatic interactions between WPI-AI led to coacervation yields >90 %, influenced by the preconditioning process employed. Viscoelastic properties outlined a predominantly solid-like behavior (G´ > G”). The CCWPI:AI system based on WPIT showed enhanced strength and gel-like structure compared to the WPIUS-based system. Oil-in-water (O/W) emulgels were formed and stabilized with the CCWPI:AI complexes, exhibiting spherical droplets (93.3–292.8 μm), whereas texture and rheological properties highlighted the formation of gel-like systems. The centrifugation STEP technology was used to evaluate the physical stability of emulgels, WPIT-based emulgels displayed superior stability against creaming than untreated WPI and WPIUS-based emulgels. These findings provide a basis for developing emulgels with prolonged stability and tunable functional properties, tailoring enhanced viscoelastic and texture attributes to meet specific needs for industrial applications where gel-like properties are pursued.

Optimizing preparation of low-NaCl protein gels from goose meat and understanding synergistic effects of pH/NaCl in improving gel characteristics

This study explored the feasibility of partially substituting NaCl with MgCl2 in preparing gel products from goose meat. Furthermore, the effects of synergistic interaction between different pH levels and NaCl concentrations on the structure and characteristics of the gels were explored by analyzing their secondary structure, microstructure, and water-distribution properties. The results showed that NaCl could be partially substituted by MgCl2, with the optimal preparation conditions: NaCl (0.83 mol/L), pH (7.3), MgCl2 (0.04 mol/L), heating temperature (79 °C), heating time (20 min), and solid-liquid ratio (1:3). Furthermore, the pH had a more significant impact on the gels' structure and characteristics than did NaCl concentration. Thus, our optimized method can reduce the usage of NaCl in the gel products while at the same time improving the characteristics of gel products under low-NaCl conditions by lowering pH, laying a solid theoretical foundation for producing low-NaCl protein gel products from goose meat.

Does particle size matter for soy protein nanoparticles in the fabrication and lipolysis of pickering-like emulsions?

Enzyme-mediated nanostructures have gained increasing interests and shown promising applications in functional delivery for food and pharmaceuticals. In our previous study, we fabricated three types of spherical soy protein nanoparticles (SPNP) by flavorzyme to different degree of hydrolysis (DH, 3%, 7% and 11%). These SPNPs exhibited similar subunit composition, secondary structure and surface properties, but differ in size, which is 89 nm, 106 nm and 151 nm, respectively. This study further applied them in emulsion systems for functional delivery. We systematically studied the emulsifying activities, interfacial properties including wettability, interfacial packing pattern, interfacial adsorption and dilatational rheological behavior of SPNPs, and their performance in modulating lipolysis of the emulsions under the influence of lipase, bile salts, pepsin and trypsin. All SPNPs behaved as Pickering-like stabilizers with moderate wettability at oil-water interface with three phase contact angle of 92–101°, and the SPNP with the smallest size has better emulsifying activities (D3,2 = 3.22 ± 0.00 μm in SDS solution) than native soy protein isolate (SPI) (D3,2 = 4.04 ± 0.03 μm in SDS solution). These SPNPs formed similar viscoelastic oil-water interfaces (Ed' = 20.2–21.5 mN/m), which were more glassy-like with weak in-plane interactions compared to SPI (Ed' = 28.6 ± 0.5 mN/m). All SPNPs can delay lipolysis of emulsions by lowering the access/replacement of lipase and bile salts and depressing the proteolysis by trypsin at interface in similar ways. Our work elucidated the role of particle size of enzyme-mediated plant protein-based particle in emulsion stabilization and lipolysis control, and is expected to give the theoretical support for the design of functional food emulsions with controllable energy intake.

The Influence of Gamma Radiation on Different Gelatin Nanofibers and Gelatins.

Gelatin nanofibers are known as wound-healing biomaterials due to their high biocompatible, biodegradable, and non-antigenic properties compared to synthetic-polymer-fabricated nanofibers. The influence of gamma radiation doses on the structure of gelatin nanofiber dressings compared to gelatin of their origin is little known, although it is very important for the production of stable bioactive products. Different-origin gelatins were extracted from bovine and donkey hides, rabbit skins, and fish scales and used for fabrication of nanofibers through electrospinning of gelatin solutions in acetic acid. Nanofibers with sizes ranging from 73.50 nm to 230.46 nm were successfully prepared, thus showing the potential of different-origin gelatin by-products valorization as a lower-cost alternative to native collagen. The gelatin nanofibers together with their origin gelatins were treated with 10, 20, and 25 kGy gamma radiation doses and investigated for their structural stability through chemiluminescence and FTIR spectroscopy. Chemiluminescence analysis showed a stable behavior of gelatin nanofibers and gelatins up to 200 °C and increased chemiluminescent emission intensities for nanofibers treated with gamma radiation, at temperatures above 200 °C, compared to irradiated gelatins and non-irradiated nanofibers and gelatins. The electron paramagnetic (EPR) signals of DMPO adduct allowed for the identification of long-life HO● radicals only for bovine and donkey gelatin nanofibers treated with a 20 kGy gamma radiation dose. Microbial contamination with aerobic microorganisms, yeasts, filamentous fungi, Staphylococcus aureus, Escherichia coli, and Candida albicans of gelatin nanofibers treated with 10 kGy gamma radiation was under the limits required for pharmaceutical and topic formulations. Minor shifts of FTIR bands were observed at irradiation, indicating the preservation of secondary structure and stable properties of different-origin gelatin nanofibers.

Tomato (Solanum lycopersicum) leaf juice induced whey protein gelling: Unveiling the potential of endogenous proteases in novel applications

Tomato leaves are by-products of tomato production and a potential source of proteins. Part of these proteins are proteases that carry out important functions for the plants. These endogenous proteases can bring challenges in downstream protein extraction, and can alter protein functionality. The aim of this study was to characterize the endogenous proteases in tomato leaf juice and to investigate the hydrolysis by these proteases on both endogenous leaf proteins and WPI. In addition, we characterized the gelation behavior, rheological properties and structural and thermal properties change of WPI by the juice proteases. 38 different proteases were identified in the juice, along with 4 protease inhibitors. The proteases hydrolyzed both endogenous leaf proteins and WPI, with predominantly an endopeptidase manner for the latter. The protein hydrolysis was positively affected by the incubation time and temperature. The protease activity in juice could not be fully eliminated by heat deactivation or the addition of protease inhibitors. The protein hydrolysis however remained limited (max. 43 %) under all conditions below 60 °C. This hydrolysis resulted in small changes in both secondary structure and thermal properties of WPI. Limited hydrolysis led to WPI gelation at lower protein concentration. Gel formed with WPI after limited hydrolysis had higher gel strength and more brittle structure than the gel with intact proteins. To conclude, the complex and active nature of proteases in tomato leaves poses challenge for downstream protein extraction. Alternatively, juice from tomato leaves could be used as natural gelling agent for WPI.

Ultrasound facilitate temperature-dependence adsorption of β-lactoglobulin on starch nanoparticles used for enhancing protein desensitization and modifying the physicochemical properties

This study aimed to probe an effective approach to reduce the allergy of β-lactoglobulin (βLg) by formation of protein corona on starch nanoparticles (SNPs) and unveil the role of ultrasound in temperature-dependence SNPs-βLg coronas systematically through the βLg adsorption mechanism, changes in secondary structure and physicochemical properties. The adsorption was found to be a spontaneous process that negatively correlate with temperature, while, sonication increased the theoretical maximum binding number of βLg from 4600 to 7800, accompanied with higher binding affinity (Ka) of 58 × 106 M−1 and greater change in secondary structure of βLg with 22 ± 3.1% of β-sheet. Functionally, ultrasound facilitated the functional properties changes to βLg induced by adsorption to SNPs that inhibited βLg's immunoglobulin E (IgE) combining capacity for 87 ± 5%, decreased surface hydrophobicity and digestibility due to the more decrease of β-sheet, whereas, increased thermal stability and emulsifying ability. These findings promoted the application potential of βLg and SNPs in food.

Effect of starch-based emulsion with different amylose content on the gel properties of Nemipterus virgatus surimi

The emulsion was prepared with peanut oil and corn starch with different amylose content using high-speed homogenization assisted high-pressure homogenization, and the effect of starch-based emulsion on the gel properties, whiteness, microstructure, protein secondary structure, chemical forces, texture and sensory properties of Nemipterus virgatus surimi was investigated. The results showed that high amylose corn starch was more beneficial to the stability of emulsion than normal and waxy starch. The gel strength, water holding capacity and texture properties of surimi were significantly improved by adding 10 % waxy corn starch-based emulsion or 15 % high amylose or normal corn starch-based emulsion. Moreover, the whiteness of surimi gel containing starch-based emulsion was higher, and the microstructure was more compact and delicate than that of surimi without emulsion. The addition of starch-based emulsion could increase the hydrophobic interaction and disulfide bond content, and promote the transformation of protein secondary structure to irregular direction. The sensory properties such as color, texture, taste and overall acceptability could be improved to varying degrees. Therefore, starch-based emulsion could be used to enhance the gel properties and nutritional value of surimi products.

The male pachynema-specific protein MAPS drives phase separation in vitro and regulates sex body formation and chromatin behaviors in vivo

Dynamic chromosome remodeling and nuclear compartmentalization take place during mammalian meiotic prophase I. We report here that the crucial roles of male pachynema-specific protein (MAPS) in pachynema progression might be mediated by its liquid-liquid phase separation invitro and in cellulo. MAPS forms distinguishable liquid phases, and deletion or mutations of its N-terminal amino acids (aa) 2-9 disrupt its secondary structure and charge properties, impeding phase separation. Maps-/- pachytene spermatocytes exhibit defects in nucleus compartmentalization, including defects in forming sex bodies, altered nucleosome composition, and disordered chromatin accessibility. MapsΔ2-9/Δ2-9 male mice expressing MAPS protein lacking aa 2-9 phenocopy Maps-/- mice. Moreover, a frameshift mutation in C3orf62, the human counterpart of Maps, is correlated with nonobstructive azoospermia in a patient exhibiting pachynema arrest in spermatocyte development. Hence, the phase separation property of MAPS seems essential for pachynema progression in mouse and human spermatocytes.

SSDraw: Software for generating comparative protein secondary structure diagrams.

The program SSDraw generates publication-quality protein secondary structure diagrams from three-dimensional protein structures. To depict relationships between secondary structure and other protein features, diagrams can be colored by conservation score, B-factor, or custom scoring. Diagrams of homologous proteins can be registered according to an input multiple sequence alignment. Linear visualization allows the user to stack registered diagrams, facilitating comparison of secondary structure and other properties among homologous proteins. SSDraw can be used to compare secondary structures of homologous proteins with both conserved and divergent folds. It can also generate one secondary structure diagram from an input protein structure of interest. The source code can be downloaded (https://github.com/ncbi/SSDraw) and run locally for rapid structure generation, while a Google Colab notebook allows easy use.

Rational design of keratin nanoparticles utilizing diverse hair protein fractions for controlled drug release

BackgroundHair keratin exhibits potential as a nanocarrier material owing to its biocompatibility, biodegradability, and adaptability. However, conventional extraction methods yield mixtures of KIFs and KAPs. While prior research has delved into keratin nanoparticles, none have specifically investigated those derived from distinct keratin subgroups. MethodsTo address this gap, we assessed the particle size, zeta potential, secondary structure, and drug release properties of spray-dried keratin nanoparticles generated from diverse KAPs/KIFs mixtures. Significant FindingsOur findings reveal that KAPs/KIFs ratios directly impact keratin nanoparticles’ protein structures and properties. When a higher proportion of KAPs is involved, there is a greater repulsive force between particles, reducing their tendency to aggregate. Conversely, increased KIFs content results in smaller particles and reduced electrostatic repulsion, potentially enhancing Theophylline release. Therefore, fine-tuning drug release behavior by adjusting KAPs and KIFs ratios while considering environmental pH values is critical to address aggregation problems while maximizing drug loading and encapsulation efficiency. These results underscore the importance of controlled keratin composition for developing reliable, keratin-based biomaterials tailored for diverse biomedical applications.

Silk-hydrogel functionalized with human decellularized Wharton's jelly extracellular matrix as a minimally invasive injectable hydrogel system for potential nucleus pulposus tissue replacement therapy

Intervertebral disc degeneration is a primary cause for chronic low back pain, a common health problem with high incidence and the leading cause of disability globally. The early stages of disc degeneration in terms of functional and anatomical abnormalities start from the central nucleus pulposus tissue of the intervertebral disc; hence its regeneration has become a prime concern. A plethora of hydrogel systems have been investigated as nucleus pulposus tissue substitute over the years, with limited clinical translation. In the present study, we formulated a minimally invasive injectable cross-linker-free bioactive silk-based hybrid hydrogel system functionalized with decellularized human Wharton's jelly extracellular matrix as an ampule of bioactive cues. The centrifugation based decellularization method removed >92 % of cellular components and preserved >83 % of extracellular matrix composition. The hydrogels were investigated for secondary structure and surface properties through infrared spectroscopy and electron micrographs, respectively. Notably, the developed hydrogels were found to mimic the rheological and mechanical properties of native nucleus pulposus tissue when decellularized Wharton's jelly extracellular matrix content was 0.5 % (w/v) in the base silk hydrogel. Finally, the hydrogels were found to support cell viability, proliferation, and tissue maturation offering great potential for future applications related to nucleus pulposus tissue engineering.

Breaking the temperature limitation of zein-rice starch dough by microwave pre-gelatinization: Morphological, structural and rheological properties of the dough

Zein has gluten-like viscoelasticity, but its use is limited due to high glass transition temperature (Tg). To break the temperature limitation of zein-starch dough, microwave heating was used to pre-gelatinize a partial of the starch with zein, and then the remaining was added and kneaded to form a dough. Pre-gelatinized doughs formed by rice starch (PRS), zein-starch (PUZS), and extruded zein-starch (PEZS) were included in this study. The thermal, morphological, rheological, and secondary structural properties of the dough were investigated. The results showed that zein and starch formed a composite gel network and firmly bound starch granules, which improved the dough properties with a smooth surface and compact internal structure, increased strain tolerance, and decreased stiffness. Unextruded zein was distributed uniformly and had strong interactions with the starch. Extruded zein tended to form large particles and had limited interaction with starch but improved dough extensibility. Microwave pre-gelatinization increased the stability of the secondary structure of zein and maintained the viscoelasticity of dough below zein’s Tg, which provided a safe and effective way to break the temperature limitation of zein as a structural protein used in foods.

Effects of different ohmic heating treatments on parvalbumin structure and reduction of allergenicity in Japanese eel (Anguilla japonica)

We investigated the effects of ohmic heating (OH) on the structural properties and allergenicity of parvalbumin (PV). Compared to other heating methods (water bath heating (WH), OH combined with WH, and OH combined with air thermostatic heating (AH)), pure OH heating expended the least time and total energy. PV sensitization was reduced by approximately 65% by pure OH heating. SDS-PAGE, tricine–SDS-PAGE, and western blotting analyses revealed a molecular weight of sensitized β-PV of about 12 kDa. Band intensity decreased with increasing OH time, and significant changes were observed in amino acid content, secondary structure, microstructure, and dielectric properties. Reducing PV, allergenicity through protein unfolding and secondary structural changes, thereby possibly reducing the allergenicity of eel, provides a theoretical basis for developing hypoallergenic products.

Isolation and Characterization of Protein Fractions for Valorization of Sacha Inchi Oil Press-Cake.

The growing interest in plant-based food protein sources has provided opportunities for the valorization of agri-food by-products, driving the food industry towards more sustainable development. In this study, three extraction procedures (varying the pH value (7.0 and 11.0) and the addition of salt (0 and 5%)) were investigated to obtain seven different protein fractions (SIPF) from Sacha Inchi oil press-cake (SIPC), which were characterized in terms of their protein content, electrophoretic profile, secondary structure, and techno-functional properties. Extractions at pH 11.0 without salt addition produced the highest values of protein content, extraction yield, protein recovery, and protein concentration increase (84.0%, 24.7%, 36.5%, and 1.5-fold, respectively). Under these extraction conditions, the electrophoretic analysis indicated that most of the SIPC proteins were extracted. SIPF displayed an excellent oil absorption capacity (4.3-9.0 w/w), and interesting foam activity (36.4-133.3%). The solubility and emulsifying activity of the albumin fractions were significantly higher than those of the other fractions (~87 vs. <15.8%, and 280-370 vs. <140 m2/g, respectively). Correlation analysis showed that the secondary structure of the SIPF significantly influences their techno-functional properties. These results indicate that SIPC is a by-product of great potential for protein extraction processes, and as a valorization strategy for technical cycle solutions for the Sacha Inchi productive chain in the circular economy context.

Insights into the material properties of dragline spider silk affecting Schwann cell migration

Dragline silk of Trichonephila spiders has attracted attention in various applications. One of the most fascinating uses of dragline silk is in nerve regeneration as a luminal filling for nerve guidance conduits. In fact, conduits filled with spider silk can measure up to autologous nerve transplantation, but the reasons behind the success of silk fibers are not yet understood.In this study dragline fibers of Trichonephila edulis were sterilized with ethanol, UV radiation, and autoclaving and the resulting material properties were characterized with regard to the silk's suitability for nerve regeneration. Rat Schwann cells (rSCs) were seeded on these silks in vitro and their migration and proliferation were investigated as an indication for the fiber's ability to support the growth of nerves. It was found that rSCs migrate faster on ethanol treated fibers. To elucidate the reasons behind this behavior, the fiber's morphology, surface chemistry, secondary protein structure, crystallinity, and mechanical properties were studied. The results demonstrate that the synergy of dragline silk's stiffness and its composition has a crucial effect on the migration of rSCs. These findings pave the way towards understanding the response of SCs to silk fibers as well as the targeted production of synthetic alternatives for regenerative medicine applications.

Amyloid fibrils degradation: the pathway to recovery or aggravation of the disease?

Background: The most obvious manifestation of amyloidoses is the accumulation of amyloid fibrils as plaques in tissues and organs, which always leads to a noticeable deterioration in the patients' condition and is the main marker of the disease. For this reason, early diagnosis of amyloidosis is difficult, and inhibition of fibrillogenesis, when mature amyloids are already accumulated in large quantities, is ineffective. A new direction for amyloidosis treatment is the development of approaches aimed at the degradation of mature amyloid fibrils. In the present work, we investigated possible consequences of amyloid's degradation. Methods: We analyzed the size and morphology of amyloid degradation products by transmission and confocal laser scanning microscopy, their secondary structure and spectral properties of aromatic amino acids, intrinsic chromophore sfGFP, and fibril-bound amyloid-specific probe thioflavin T (ThT) by the absorption, fluorescence and circular dichroism spectroscopy, as well as the cytotoxicity of the formed protein aggregates by MTT-test and their resistance to ionic detergents and boiling by SDS-PAGE. Results: On the example of sfGFP fibrils (model fibrils, structural rearrangements of which can be detected by a specific change in the spectral properties of their chromophore), and pathological Aβ-peptide (Aβ42) fibrils, leading to neuronal death in Alzheimer's disease, the possible mechanisms of amyloids degradation after exposure to factors of different nature (proteins with chaperone and protease activity, denaturant, and ultrasound) was demonstrated. Our study shows that, regardless of the method of fibril degradation, the resulting species retain some amyloid's properties, including cytotoxicity, which may even be higher than that of intact amyloids. Conclusion: The results of our work indicate that the degradation of amyloid fibrils in vivo should be treated with caution since such an approach can lead not to recovery, but to aggravation of the disease.