Fibril Structure Research Articles

Oleogels based on peanut protein isolate fibrils: Structural characterization dependent on induction time and suitability in marguerite biscuits

In order to develop new plant protein-based lipid substitutes to deal with health problems caused by animal fats or trans-fatty acids, the emulsion stablized by acid-heat (90 °C, pH 2.0) induced aggregation of peanut protein isolate (PPI) was used as template to fabricate oleogels. Heating time affected the structure, morphology of PPI and the adsorption characteristics at the O/W interface. The protein with suitable heating time (2–6 h) formed fibrils structure, which was beneficial to the rapid adsorption of aggregates at the O/W interface. Overheating (exceeding 8 h) led to the aggregate of protein fibrils and inhibited the adsorption of proteins at the O/W interface. Therefore, the heating times of 2 h, 4 h and 6 h were selected to prepare the follow-up emulsion and oleogels. Among the four heating times, the emulsion prepared by PPI heating for 6 h showed the best storage stability, ionic strength stability and shear resistance. The resultant oleogels exhibited the least oil loss and the highest freeze-thaw stability. The margarita biscuits prepared with a substitution rate of 50% oleogels to butter presented similar sensory property to the margaritas prepared with butter, indicating that the oleogels possessed favorable shortening properties. The results indicated that PPI fibrils structure could improve the oleogel formaton properties, making it a potential novel additive for the development of new fat substitute products.

An electrostatic cluster guides Aβ40 fibril formation in sporadic and Dutch-type cerebral amyloid angiopathy

Cerebral amyloid angiopathy (CAA) is associated with the accumulation of fibrillar Aβ peptides upon and within the cerebral vasculature, which leads to loss of vascular integrity and contributes to disease progression in Alzheimer’s disease (AD). We investigate the structure of human-derived Aβ40 fibrils obtained from patients diagnosed with sporadic or familial Dutch-type (E22Q) CAA. Using cryo-EM, two primary structures are identified containing elements that have not been observed in in vitro Aβ40 fibril structures. One population has an ordered N-terminal fold comprised of two β-strands stabilized by electrostatic interactions involving D1, E22, D23 and K28. This charged cluster is disrupted in the second population, which exhibits a disordered N-terminus and is favored in fibrils derived from the familial Dutch-type CAA patient. These results illustrate differences between human-derived CAA and AD fibrils, and how familial CAA mutations can guide fibril formation.

Rational design of structure-based vaccines targeting misfolded alpha-synuclein conformers of Parkinson's disease and related disorders.

Synucleinopathies, including Parkinson's disease (PD), multiple system atrophy (MSA), and dementia with Lewy bodies (DLB), are neurodegenerative disorders caused by the accumulation of misfolded alpha-synuclein protein. Developing effective vaccines against synucleinopathies is challenging due to the difficulty of stimulating an immune-specific response against alpha-synuclein without causing harmful autoimmune reactions, selectively targeting only pathological forms of alpha-synuclein. Previous attempts using linear peptides and epitopes without control of the antigen structure failed in clinical trials. The immune system was unable to distinguish between native alpha-synuclein and its amyloid form. The prion domain of the fungal HET-s protein was selected as a scaffold to introduce select epitopes from the surface of alpha-synuclein fibrils. Four vaccine candidates were generated by introducing specific amino acid substitutions onto the surface of the scaffold protein. The approach successfully mimicked the stacking of the parallel in-register beta-sheet structure seen in alpha-synuclein fibrils. All vaccine candidates induced substantial levels of IgG antibodies that recognized pathological alpha-synuclein fibrils derived from a synucleinopathy mouse model. Furthermore, the antisera recognized pathological alpha-synuclein aggregates in brain lysates from patients who died from DLB, MSA, or PD, but did not recognize linear alpha-synuclein peptides. Our approach, based on the rational design of vaccines using the structure of alpha-synuclein amyloid fibrils and strict control over the exposed antigen structure used for immunization, as well as the ability to mimic aggregated alpha-synuclein, provides a promising avenue toward developing effective vaccines against alpha-synuclein fibrils.

A Model Study to Assess Fibrillation and Product Stability to Support Peptide Drug Design.

The fibrillation of therapeutic peptides can present significant quality concerns and poses challenges for manufacturing and storage. A fundamental understanding of the mechanisms of fibrillation is critical for the rational design of fibrillation-resistant peptide drugs and can accelerate product development by guiding the selection of solution-stable candidates and formulations. The studies reported here investigated the effects of structural modifications on the fibrillation of a 29-residue peptide (PepA) and two sequence modified variants (PepB, PepC). The C-terminus of PepA was amidated, whereas both PepB and PepC retained the carboxylate, and Ser16 in PepA and PepB was substituted with a helix-stabilizing residue, α-aminoisobutyric acid (Aib), in PepC. In thermal denaturation studies by far-UV CD spectroscopy and fibrillation kinetic studies by fluorescence and turbidity measurements, PepA and PepB showed heat-induced conformational changes and were found to form fibrils, whereas PepC did not fibrillate and showed only minor changes in the CD signal. Pulsed hydrogen-deuterium exchange mass spectrometry (HDX-MS) showed a high degree of protection from HD exchange in mature PepA fibrils and its proteolytic fragments, indicating that most of the sequence had been incorporated into the fibril structure and occurred nearly simultaneously throughout the sequence. The effects of the net peptide charge and formulation pH on fibrillation kinetics were investigated. In real-time stability studies of two formulations of PepA at pH's 7.4 and 8.0, analytical methods detected significant changes in the stability of the formulations at different time points during the study, which were not observed during accelerated studies. Additionally, PepA samples were withdrawn from real-time stability and subjected to additional stress (40 °C, continuous shaking) to induce fibrillation; an approach that successfully amplified oligomers or prefibrillar species previously undetected in a thioflavin T assay. Taken together, these studies present an approach to differentiate and characterize fibrillation risk in structurally related peptides under accelerated and real-time conditions, providing a model for rapid, iterative structural design to optimize the stability of therapeutic peptides.

Structure of alpha-synuclein fibrils derived from human Lewy body dementia tissue

The defining feature of Parkinson disease (PD) and Lewy body dementia (LBD) is the accumulation of alpha-synuclein (Asyn) fibrils in Lewy bodies and Lewy neurites. Here we develop and validate a method to amplify Asyn fibrils extracted from LBD postmortem tissue samples and use solid state nuclear magnetic resonance (SSNMR) studies to determine atomic resolution structure. Amplified LBD Asyn fibrils comprise a mixture of single protofilament and two protofilament fibrils with very low twist. The protofilament fold is highly similar to the fold determined by a recent cryo-electron microscopy study for a minority population of twisted single protofilament fibrils extracted from LBD tissue. These results expand the structural characterization of LBD Asyn fibrils and approaches for studying disease mechanisms, imaging agents and therapeutics targeting Asyn.

Phosphorylation and O-GlcNAcylation at the same α-synuclein site generate distinct fibril structures

α-Synuclein forms amyloid fibrils that are critical in the progression of Parkinson’s disease and serves as the pathological hallmark of this condition. Different posttranslational modifications have been identified at multiple sites of α-synuclein, influencing its conformation, aggregation and function. Here, we investigate how disease-related phosphorylation and O-GlcNAcylation at the same α-synuclein site (S87) affect fibril structure and neuropathology. Using semi-synthesis, we obtained homogenous α-synuclein monomer with site-specific phosphorylation (pS87) and O-GlcNAcylation (gS87) at S87, respectively. Cryo-EM revealed that pS87 and gS87 α-synuclein form two distinct fibril structures. The GlcNAc situated at S87 establishes interactions with K80 and E61, inducing a unique iron-like fold with the GlcNAc molecule on the iron handle. Phosphorylation at the same site prevents a lengthy C-terminal region including residues 73 to 140 from incorporating into the fibril core due to electrostatic repulsion. Instead, the N-terminal half of the fibril (1–72) takes on an arch-like fibril structure. We further show that both pS87 and gS87 α-synuclein fibrils display reduced neurotoxicity and propagation activity compared with unmodified α-synuclein fibrils. Our findings demonstrate that different posttranslational modifications at the same site can produce distinct fibril structures, which emphasizes link between posttranslational modifications and amyloid fibril formation and pathology.

Self-Assembling Gelatin-Curdlan Fibril Hydrogels for Oriented Neural Cell Growth.

The ex vivo replication of the highly helical and fibril structures of load-bearing soft tissue is a challenging goal for the study of hydrogels. Inspired by nature, we prepared tissue-like physical gels based on curdlan and gelatin by self-assembly. The hybrid gels have a flexible fibril-matrix architecture, and the fibril orientation is highly tunable. The tensile strength of the gels can be tuned from ∼1.1 to ∼16.5 MPa. The coil-helix transition and nanofibril formation process in the self-assembly system was thoroughly investigated. These helical gels exhibit excellent cell compatibility, which supports adhesion and oriented growth of neural cells. Furthermore, the oriented nanofibrils in the gel are found to be associated with an upregulated expression of regeneration-related genes like N-cadherin (Cdh2) and neural growth factor (NGF). Owing to the strength and biomimetic structure, these gels have great potential in tissue engineering applications.

Structural insight into Escherichia coli CsgA amyloid fibril assembly.

The discovery of functional amyloids in bacteria dates back several decades, and our understanding of the Escherichia coli curli biogenesis system has gradually expanded over time. However, due to its high aggregation propensity and intrinsically disordered nature, CsgA, the main structural component of curli fibrils, has eluded comprehensive structural characterization. Recent advancements in cryo-electron microscopy (cryo-EM) offer a promising tool to achieve high-resolution structural insights into E. coli CsgA fibrils. In this study, we outline an approach to addressing the colloidal instability challenges associated with CsgA, achieved through engineering and electrostatic repulsion. Then, we present the cryo-EM structure of CsgA fibrils at 3.62 Å resolution. This structure provides new insights into the cross-β structure of E. coli CsgA. Additionally, our study identifies two distinct spatial arrangements within several CsgA fibrils, a 2-CsgA-fibril pair and a 3-CsgA-fibril bundle, shedding light on the intricate hierarchy of the biofilm extracellular matrix and laying the foundation for precise manipulation of CsgA-derived biomaterials.IMPORTANCEThe visualization of the architecture of Escherichia coli CsgA amyloid fibril has been a longstanding research question, for which a high-resolution structure is still unavailable. CsgA serves as a major subunit of curli, the primary component of the extracellular matrix generated by bacteria. The support provided by this extracellular matrix enables bacterial biofilms to resist antibiotic treatment, significantly impacting human health. CsgA has been identified in members of Enterobacteriaceae, with pathogenic E. coli being the most well-known model system. Our novel insights into the structure of E. coli CsgA protofilaments form the basis for drug design targeting diseases associated with biofilms. Additionally, CsgA is widely researched in biomaterials due to its self-assembly characteristics. The resolved spatial arrangements of CsgA amyloids revealed in our study will further enhance the precision design of functional biomaterials. Therefore, our study uniquely contributes to the understanding of CsgA amyloids for both microbiology and material science.

Plant oil fillers toughened poly(3-hydroxybutyrate) green biocomposites

Poly(3-hydroxybutyrate) (PHB) faces a significant challenge of intrinsic brittleness, thus limiting its widespread applicability. This challenge can be surmounted by incorporating various nanofillers into the PHB matrix to tune the mechanical performance. However, many of these fillers are produced using unsustainable, non-environmentally friendly methods. Herein, we employed a green process to develop castor-oil-based fillers, CO-g-DLA, and CO-P(CL-co-DLA)-PDLA (CO-r-DLA), for the toughening of the PHB matrix. Impressively, the addition of 20% of these fillers increases the elongation at the break by 13-fold through forming elongated fibril structures and enhanced interfacial interactions between fillers and the matrix, accompanying by a substantial reduction on the PHB’s brittleness through shear yielding and fibrillation. These are confirmed by Small-Angle X-ray Scattering (SAXS) results, showing significant copolymer-induced microstructural changes in the matrix. The biodegradable PHB biocomposite developed in this study offers a promising green alternative to conventional plastics, especially in sustainable packaging.

Post-translational modification sites are present in hydrophilic cavities of alpha-synuclein, tau, FUS, and TDP-43 fibrils: A molecular dynamics study.

Hydration plays a crucial role in the refolding of intrinsically disordered proteins into amyloid fibrils; however, the specific interactions between water and protein that may contribute to this process are still unknown. In our previous studies of alpha-synuclein (aSyn), we have shown that waters confined in fibril cavities are stabilizing features of this pathological fold; and that amino acids that hydrogen bond with these confined waters modulate primary and seeded aggregation. Here, we extend our aSyn molecular dynamics (MD) simulations with three new polymorphs and correlate MD trajectory information with known post-translational modifications (PTMs) and experimental data. We show that cavity residues are more evolutionarily conserved than non-cavity residues and are enriched with PTM sites. As expected, the confinement within hydrophilic cavities results in more stably hydrated amino acids. Interestingly, cavity PTM sites display the longest protein-water hydrogen bond lifetimes, three-fold greater than non-PTM cavity sites. Utilizing the deep mutational screen dataset by Newberry et al. and the Thioflavin T aggregation review by Pancoe et al. parsed using a fibril cavity/non-cavity definition, we show that hydrophobic changes to amino acids in cavities have a larger effect on fitness and aggregation rate than residues outside cavities, supporting our hypothesis that these sites are involved in the inhibition of aSyn toxic fibrillization. Finally, we expand our study to include analysis of fibril structures of tau, FUS, TDP-43, prion, and hnRNPA1; all of which contained hydrated cavities, with tau, FUS, and TDP-43 recapitulating our PTM results in aSyn fibril cavities.

Modulation of Primary and Secondary Processes in Tau Fibril Formation by Salt-Induced Dynamics.

The initial stages of amyloid fibrilization begin with the monomers populating aggregation-prone conformers. Characterization of such aggregation-prone conformers is crucial in the study of neurodegenerative diseases. The current study characterizes the aggregation pathway of two tau protein constructs that have been recently demonstrated to form Alzheimer's (AD) fibril structures with divalent ions and chronic traumatic encephalopathy (CTE) fibril structures with monovalent ions. The results highlight the involvement of identical residues in both the primary and secondary processes of both AD and CTE fibril propagation. Nuclear magnetic resonance relaxation experiments reveal increased flexibility of the motifs 321KCGS within R3 and 364PGGGN within R4 in the presence of MgCl2/NaCl, correlating with faster aggregation kinetics and indicating efficient primary nucleation. Notably, the seeded aggregation kinetics of the tau monomers in the presence and absence of metal ions are strikingly different. This correlates with the overall sign of the 15N-ΔR2 profile specifying the dominant mechanism involved in the process of aggregation.

Length and saturation of choline plasmalogens alter the aggregation rate of α-synuclein but not the toxicity of amyloid fibrils

Plasmalogens comprise a large fraction of the total phospholipids in plasma membranes. These molecules modulate membrane fluidity, produce inflammatory mediators mitigating effects of metabolic stresses. A growing body of evidence suggests that an onset of Parkinson's disease (PD), a severe neurodegenerative pathology, can be triggered by metabolic changes in plasma membranes. However, the role of plasmalogens in the aggregation of α-synuclein (α-syn), an expected molecular cause of PD, remains unclear. In this study we examine the effect of choline plasmalogens (CPs), unique phospholipids that have a vinyl ether linkage at the sn-1 position of glycerol, on the aggregation rate of α-syn. We found that the length and saturation of fatty acids (FAs) in CPs change rates of protein aggregation. We also found drastic changes in the morphology of α-syn fibrils formed in the presence of different CPs compared to α-syn fibrils grown in the lipid-free environment. At the same time, we did not observe substantial changes in the secondary structure and toxicity of α-syn fibrils formed in the presence of different CPs. These results indicate that the length and saturation of FAs in CPs present in the plasma membrane can alter α-syn stability and modulate its aggregation properties, which, in turn can accelerate or delay the onset of PD.

Abstract 1610 Leveraging the structures of transthyretin fibrils to understand and detect protein aggregation in ATTR amyloidosis

Abstract 1610 Leveraging the structures of transthyretin fibrils to understand and detect protein aggregation in ATTR amyloidosis

Review: fish collagen: extraction, characterization and application in wound healing and drug delivery

Collagen is a natural protein presenting in the animal and human’s body. It is in natural fibrous and could be found in the connective tissue and flesh of mammal, approximately 25-35 wt.% of total human protein. Collagen is abundant in fibrous tissues such as: bones, cartilage, tendons, blood vessels, ligaments, skin, cornea, aortic disc and intestines. Fish collagen is currently being studied as a new biological substitute for collagen from animals. It extracted from by-products (scales, skins, bones, etc.) by biochemical or biological methods is safe, has high adsorption, high compatible. The factors influencing on the extraction process include temperature, time, concentration of ingredients, etc. The fish collagen is usually collagen type I with a fibril structure. Each unit of collagen is tropocollagen consisting of three polypeptide chains twisted together. Depending on the source of ingredients, the composition of aminoacids, denaturation temperature, the sequence of aminoacids, structure, morphology, molecular weight, extraction yield, etc. of obtained collagens are different. This paper reviews the extraction and characterization of fish collagen from products after fish processing by chemical or biochemical methods. The applications of fish collagen in wound healing and drug delivery are also reviewed.

Linear sensing actuators of polypyrrole nanofiber scaffolds

AbstractIn the search for renewable materials for linear actuators, polypyrrole (PPy) glucose‐glycerin nanofiber scaffolds were used here to provide the actuator with a basic fibrillary structure, mimicking natural muscles. Each nanofiber was coated with a chemically synthesized PPy film, getting conductive nanofiber scaffolds (CNS), used then as electrodes to attain a second polypyrrole doped with dodecylbenzensulfonate (PPyDBS) coat forming PPyDBS‐CNS material structure. PPyDBS bulk films obtained by electropolymerization on stainless steel under the same condition were used to compare the linear actuation properties of both materials. Cyclic voltammetry, square potential waves, and square potential currents, in combination with linear actuation measurements, studied the samples. Three different potential ranges (PRs) were selected for those methodologies: 1.0 to −0.55 V (PR1), 0.8 to −0.4 V (PR2), and 0.65 to −0.55 V (PR3), revealing that PPyDBS‐CNS has anion‐driven actuation independent of the applied PR1–3, while in comparison PPyDBS films had in PR3 mixed ion actuation. The best strain from PPyDBS, 24.6%, was attained at PR1, and from PPyDBS‐CSN, 17.5% strain in the same PR. Further characterizations are conducted, such as scanning electron microscopy, Fourier transform infrared spectroscopy, and element determination using energy dispersive x‐ray spectroscopy.

Edge Substitution Effects of Histidine Tautomerization Behaviors on the Structural Properties and Aggregation Properties of Aβ(1-42) Mature Fibril.

Histidine behaviors play critical roles in folding and misfolding processes due to the changes in net charge and the various N/N-H orientations on imidazole rings. However, the effect of histidine tautomerization (HIE (Nε-H, ε) and HID (Nδ-H, δ) states) behaviors on the edge chain of Aβ mature fibrils remains inadequately understood, which is critical for finding a strategy to disturb fibril elongation and growth. In the current study, eight independent molecular dynamics simulations were conducted to investigate such impacts on the structural and aggregation properties. Our results from three different binding models revealed that the binding contributions of edge substitution effects are primarily located between chains 1 and 2. Histidine states significantly influence the secondary structure of each domain. Further analysis confirmed that the C1_H6//C1_E11 intrachain interaction is essential in maintaining the internal stability of chain 1, while the C1_H13//C2_H13 and C1_H14//C2_H13 interchain interactions are critical in maintaining the interchain stability of the fibril structure. Our subsequent analysis revealed that the current edge substitution leads to the loss of the C1_H13//C1_E11 intrachain and C1_H13//C2_H14 interchain interactions. The N-terminal regularity was significantly directly influenced by histidine states, particularly by the residue of C1_H13. Our study provides valuable insights into the effect of histidine behaviors on the edge chain of Aβ mature fibril, advancing our understanding of the histidine behavior hypothesis in misfolding diseases.

Large Unilamellar Vesicles of Phosphatidic Acid Reduce the Toxicity of α-Synuclein Fibrils.

Parkinson's disease (PD) is a severe pathology that is caused by a progressive degeneration of dopaminergic neurons in substantia nigra pars compacta as well as other areas in the brain. These neurodegeneration processes are linked to the abrupt aggregation of α-synuclein (α-syn), a small protein that is abundant at presynaptic nerve termini, where it regulates cell vesicle trafficking. Due to the direct interactions of α-syn with cell membranes, a substantial amount of work was done over the past decade to understand the role of lipids in α-syn aggregation. However, the role of phosphatidic acid (PA), a negatively charged phospholipid with a small polar head, remains unclear. In this study, we examined the effect of PA large unilamellar vesicles (LUVs) on α-syn aggregation. We found that PA LUVs with 16:0, 18:0, and 18:1 FAs drastically reduced the toxicity of α-syn fibrils if were present in a 1:1 molar ratio with the protein. Our results also showed that the presence of these vehicles changed the rate of α-syn aggregation and altered the morphology and secondary structure of α-syn fibrils. These results indicate that PA LUVs can be used as a potential therapeutic strategy to reduce the toxicity of α-syn fibrils formed upon PD.

Improving cryo-EM grids for amyloid fibrils using interface-active solutions and spectator proteins

Preparation of cryoelectron microscopy (cryo-EM) grids for imaging of amyloid fibrils is notoriously challenging. The human islet amyloid polypeptide (hIAPP) serves as a notable example, as the majority of reported structures have relied on the use of nonphysiological pH buffers, N-terminal tags, and seeding. This highlights the need for more efficient, reproducible methodologies that can elucidate amyloid fibril structures formed under diverse conditions. In this work, we demonstrate that the distribution of fibrils on cryo-EM grids is predominantly determined by the solution composition, which is critical for the stability of thin vitreous ice films. We discover that, among physiological pH buffers, HEPES uniquely enhances the distribution of fibrils on cryo-EM grids and improves the stability of ice layers. This improvement is attributed to direct interactions between HEPES molecules and hIAPP, effectively minimizing the tendency of hIAPP to form dense clusters in solutions and preventing ice nucleation. Furthermore, we provide additional support for the idea that denatured protein monolayers forming at the interface are also capable of eliciting a surfactant-like effect, leading to improved particle coverage. This phenomenon is illustrated by the addition of nonamyloidogenic rat IAPP (rIAPP) to a solution of preaggregated hIAPP just before the freezing process. The resultant grids, supplemented with this “spectator protein”, exhibit notably enhanced coverage and improved ice quality. Unlike conventional surfactants, rIAPP is additionally capable of disentangling the dense clusters formed by hIAPP. By applying the proposed strategies, we have resolved the structure of the dominant hIAPP polymorph, formed in vitro at pH 7.4, to a final resolution of 4 Å. The advances in grid preparation presented in this work hold significant promise for enabling structural determination of amyloid proteins which are particularly resistant to conventional grid preparation techniques.

Roles of soybean β-conglycinin subunit fractions in fibril formation and the effects of glycinin on them

This research is aimed at exploring the fibrillation of soybean β-conglycinin (7S) subunits (αα′ and β) at 90 °C and pH 2.0, as well as the effects of glycinin (11S) on them. Fibril formation kinetics was monitored by measuring changes in thioflavin T (Th T) fluorescence intensity of the proteins. This analysis showed that αα′ subunits had a higher fibrillation capacity but slower fibril formation rate than the β subunit. Glycinin accelerated the rate of fibrillation of the αα′ and β subunits but weakened the fibril structure. The size of the fibrils first decreased and then increased with increasing heating time. Circular dichroism and intrinsic fluorescence spectroscopy analysis indicated that the β-sheet structure and hydrophobic interactions were important for the generation of ordered fibrils. Shear viscosity and transmission electron microscopy analysis suggested that the αα′ subunits formed relatively rigid fibrils that led to a high solution viscosity. In contrast, β subunit fibrils became unstable after prolonged heating, forming shorter fibrils (fragments) that tended to aggregate, which led to a lower solution viscosity. This research also suggested that the extension region of αα′ subunits played an important role in fibrillation, and the presence of the highly hydrophobic basic polypeptides of glycinin might be the main reason for the inhibition of soy protein fibrillation. Our analysis of soy protein fibrillization at the subunit level provides insights that might enhance the future application of soy protein fibrils as functional ingredients in foods.

Cryo-EM structures of lipidic fibrils of amyloid-β (1-40)

Alzheimer’s disease (AD) is a progressive and incurable neurodegenerative disease characterized by the extracellular deposition of amyloid plaques. Investigation into the composition of these plaques revealed a high amount of amyloid-β (Aβ) fibrils and a high concentration of lipids, suggesting that fibril-lipid interactions may also be relevant for the pathogenesis of AD. Therefore, we grew Aβ40 fibrils in the presence of lipid vesicles and determined their structure by cryo-electron microscopy (cryo-EM) to high resolution. The fold of the major polymorph is similar to the structure of brain-seeded fibrils reported previously. The majority of the lipids are bound to the fibrils, as we show by cryo-EM and NMR spectroscopy. This apparent lipid extraction from vesicles observed here in vitro provides structural insights into potentially disease-relevant fibril-lipid interactions.