Hydrophilic Cavity Research Articles

Programmed polymersomes with spatio-temporal delivery of antigen and dual-adjuvants for efficient dendritic cells-based cancer immunotherapy

Since antigen and adjuvant are rapid clearance in vivo, insufficient delivery to induce dendritic cells (DCs) maturation and cross-presentation, as well as limited migration efficiency of DCs to secondary lymph organs, greatly hinders the development of DCs-based immunotherapy. Herein, PCL-PEG-PCL polymersomes (PCEP-PS) as antigen and adjuvants delivery nanoplatforms (IMO-PS) were well-designed, which can electrostatically adsorb OVA antigen on the surface via DOTAP lipid and effectively encapsulate OVA antigen into the inner hydrophilic cavity to achieve both initial antigen exposure as well as slow and sustained antigen release, incorporate MPLA within the lipid layer to ligate with extracellular TLR4 of DCs as well as encapsulate IMQ in the hydrophobic membrane to ligate with intracellular TLR7/8 of DCs for activating synergistic immune responses via different signaling pathways. The IMO-PS significantly improved antigen uptake, promoted DCs maturation and cytokines production. DCs treated with IMO-PS could enhance migration into draining lymphoid nodes, and eventually induced antigen-specific CD8+ and CD4+ T cell responses and OVA-specific cytotoxic T lymphocyte (CTL) responses. Prophylactic vaccination of EG7-OVA tumor-bearing mice by IMO-PS + DCs significantly extended tumor-free time, effectively suppressed tumor growth, and greatly extended median survival time. The strategy may provide an effective nanoplatform for co-delivery antigen and dual-adjuvants in a spatio-temporally programmed manner for DC-based cancer immunotherapy.

Structural characterization of proton-pumping rhodopsin lacking a cytoplasmic proton donor residue by X-ray crystallography

DTG/DTS rhodopsin, which was named based on a three-residue motif (DTG or DTS) that is important for its function, is a light-driven proton-pumping microbial rhodopsin using a retinal chromophore. In contrast to other light-driven ion-pumping rhodopsins, DTG/DTS rhodopsin does not have a cytoplasmic proton donor residue, such as Asp, Glu, or Lys. Because of the lack of cytoplasmic proton donor residue, proton directly binds to the retinal chromophore from the cytoplasmic solvent. However, mutational experiments that showed the complicated effects of mutations were not able to clarify the roles played by each residue, and the detail of proton uptake pathway is unclear because of the lack of structural information. To understand the proton transport mechanism of DTG/DTS rhodopsin, here we report the three-dimensional structure of one of the DTG/DTS rhodopsins, PspR from Pseudomonas putida, by X-ray crystallography. We show that the structure of the cytoplasmic side of the protein is significantly different from that of bacteriorhodopsin, the best-characterized proton-pumping rhodopsin, and large cytoplasmic cavities were observed. We propose that these hydrophilic cytoplasmic cavities enable direct proton uptake from the cytoplasmic solvent without the need for a specialized cytoplasmic donor residue. The introduction of carboxylic residues homologous to the cytoplasmic donors in other proton-pumping rhodopsins resulted in higher pumping activity with less pH dependence, suggesting that DTG/DTS rhodopsins are advantageous for producing energy and avoiding intracellular alkalization in soil and plant-associated bacteria.

Extra-thin composite nanofiltration membranes tuned by γ-cyclodextrins containing amphipathic cavities for efficient separation of magnesium/lithium ions

Although nanofiltration (NF) membrane as a new frontier technique had made significant progress in the field of lithium extraction from salt lakes, they still suffered from the conventional permeability–selectivity trade-off effect. In this study, γ-cyclodextrins(CDs) containing amphipathic cavities and polyethyleneimine (PEI) acted as the aqueous phase monomer, which was carried out interfacial polymerization with the trimesoyl chloride (TMC) as the organic phase monomer to fabricate thin film composite (TFC) NF membranes with the extra-thin active layer. The hydrophilic surface and hydrophobic cavity of γ-CDs enhanced the hydrophilicity of the membrane surface and the water flux of the nanofiltration membrane. Moreover, the cavity size of γ-CDs also effectively screened magnesium and lithium ions to a certain extent. Under the best preparation conditions, the permeation flux of the PEI/γ-CDs-TMC nanofiltration membrane was 4.86 L⋅m−2⋅h−1⋅bar−1, SLi, Mg was 10.8. It was noteworthy that we used the molecular dynamics simulation method to confirm that the participation of γ-CDs led to the diffusion rate of the water phase monomers during the interfacial polymerization reaction slowed down. Thus, the thickness of the active layer of the nanofiltration membrane formed was also significantly reduced from 110.16 nm to 52.84 nm. This PEI/γ-CDs-TMC membrane provided a new strategy for preparing nanofiltration membranes with excellent overall magnesium-lithium separation performance.

Experimental study on interaction between wettability configuration and dewing efficiency of surfaces with a cavity array

Factors that dominate dew productivity of a hydrophobic surface with a spherical hydrophilic cavity array are investigated.

Highly efficient potentiometric sensing device for gadolinium based on Tetraazacyclododecane-1, 4, 7, 10 -tetraaceticacid crown ether and multiwalled carbon nanotube composite

A hydrophilic cavity presents in tetraazacyclododecane-1, 4, 7, 10-tetraaceticacid crown ether (DOTA) provides the opportunity for Gadolinium (Gd3+) transfer from an aqueous medium to a lipophilic phase and mimic the ionophoric properties due to strong coordinating link. Therefore, we have developed Gd3+ ion selective electrode (ISE) incorporating synthesized nanocomposite of DOTA as an ionophore and multiwalled carbon nanotube (MWCNT) as signal enhancer in poly vinyl chloride (PVC) membrane sensor employing potentiometry. The composite DOTA-MWCNT was characterized by scanning electron microscopy (SEM). The Nernstian slope of 20.05 ± 0.18 mV per decade with linear range 1 × 10-2 to 1 × 10-8 M and the detection limit was found to be 7.2 ± 0.16 × 10-9 M. This electrode has an excellent selectivity over mono, bi, tri and tetravalent interferent metal species. The proposed sensor was favourably applied as an indicator electrode in the titration of Gd3+ ions with EDTA and recovery of gadolinium (III) from diverse synthetic samples. The presence of four ionizable carboxylic acids groups imparts advantages to DOTA-MWCNT composite in the formation of strong complexes and enhanced solubility of Gd3+ which became the basis for in-vivo analysis of Gd3+ in pharmaceutical and soil samples. DOTA-MWCNT as host forms 1:1 chelate complex with guest Gd3+ employing square wave voltammetry. The stability constant (Log K = 25.41) and Gibbs free energy (DG) of DOTA-MWCNT - Gd3+ complex was calculated which established exothermic and spontaneous inclusion mechanism. The higher stability constant obtained for DOTA-MWCNT chelate complex can provide the opportunity to researchers of biomedical science to minimize/control the amount of free Gd3+ release in the human body through the process of drug delivery.

Study of a water-soluble supramolecular complex of curcumin and β-cyclodextrin polymer with electrochemical property and potential anti-cancer activity

As a member of the curcuminoid compound family, curcumin (Cur) has many interesting therapeutic properties. However, its low aqueous solubility and stability have resulted in poor bioavailability and restricted clinical efficacy. Based on size matching, β-cyclodextrin polymer (β-CDP), with its hydrophilic polymer chains and hydrophobic cavities, can form an inclusion complex with Cur. To improve the water solubility and stability of Cur, a simple and eco-friendly grinding method was designed to form β-CDP inclusion complexes. According to the Boltzmann–Hamel's method and Job's method, the molar ratio of the β-CD unit in β-CDP to Cur was determined to be 1:1. The diffusion coefficient and diffusion activation energy of Cur-β-CDP were calculated in an electrochemical study. This supramolecular complex worked well in vitro to inhibit the proliferation of hepatoma carcinoma cells HepG2. Remarkably, this method visibly reduced the undesirable side effects on normal cells, without weakening the anti-cancer activity of the drugs. We expect that the obtained host–guest complex will provide a new approach for delivering natural drug molecules, having low water solubility.

Tumor microcalcification-mediated relay drug delivery for photodynamic immunotherapy of breast cancer

Spatiotemporal targeting of tumor-associated macrophages (TAMs) and tumor cells is emerging as a promising strategy for tumor therapy. Tumor microcalcifications that specifically bind to bisphosphonates are potentially used to design efficient relay drug delivery nanosystems to achieve spatiotemporal drug modulation. Here, we developed manganese dioxide (MnO2)-embedded and LyP-1 peptide-labeled liposomal nanoparticles (NPs) for photodynamic immunotherapy of breast cancer; zoledronic acid (Zol) was encapsulated in the hydrophilic cavity of liposomes, and a hydrophobic photosensitizer (IR780) was embedded in the phospholipid bilayer of liposomes. These Lipo Zol/IR NPs generated O2 bubbles through MnO2 in response to H2O2 in the tumor microenvironment, leading to the degradation of the liposomal membrane, which triggered the release of Zol and provided O2 for photodynamic therapy. The released Zol attached to microcalcifications and was selectively phagocytosed by TAMs, leading to the induction of death or repolarization of TAMs from the immunosuppressive M2 phenotype to the immunostimulatory M1 phenotype. The remaining liposomal fragments embedded with IR780 then preferentially targeted tumor cells through LyP-1 peptide and produced abundant reactive oxygen species (ROS) under near infrared (NIR) laser irradiation, resulting in the death of tumor cells and mild immune activation. All in vitro and in vivo studies demonstrated the effective photodynamic and immunoregulatory performance of Lipo Zol/IR NPs. Statement of significanceSpatiotemporal targeting of tumor-associated macrophages (TAMs) and tumor cells remains a challenge in tumor photodynamic immunotherapy for promoting synergy and reducing side effects. Here, we developed tumor microcalcification-mediated relay drug delivery nanoliposomes for breast cancer therapy. H2O2 in the tumor microenvironment (TME) triggers the breakage of nanoliposomes, thereby causing the separation of zoledronic acid (Zol) and the photosensitizer IR780 and allowing them to perform their respective functions. Microcalcifications enable Zol to target TAMs, resulting in immunomodulation. LyP-1 guides IR780 to target tumor cells for PDT with adequate O2 supply. These nanoliposomes enable precise spatiotemporal targeting of different types of cells in the TME and promote the synergy between immunotherapy and PDT while ensuring the effectiveness of both methods.

Vesicular IFN-γ as a cooperative attacker to enhance anti-cancer effect of 5-fluorouracil via thymidine phosphorylase upregulation and tumor microenvironment normalization

On the basis of immuno-modulating effect and upregulating the activity of thymidine phosphorylase (TP), interferon-γ (IFN-γ) as a cooperative attacker was explored to enhance the anticancer activity of 5-fluorouracil (5-FU). We designed and prepared a self-assembled nano-vesicular system IFN-γ-EDP formulated by amphiphilic poly((polyethylene glycol)(dodecylphosphoethanolamine)phosphazene) (EDP) to entrap IFN-γ in the hydrophilic cavity. The IFN-γ-EDP vesicles allowed IFN-γ to accumulate at the tumor site and be taken up by tumor cells, resulting in significantly upregulated expression level of TP, distinct inhibition of cell growth, more cellular apoptosis and more serious cell cycle arrest when administrated combined with 5-FU. Moreover, IFN-γ-EDP could normalize the tumor microenvironment by enhancing the CD4+ and CD8+ T cell populations, promoting the IL-12 secretion and suppressing the IL-10 secretion in tumor. As a consequence, the combination therapy of IFN-γ-EDP with 5-FU achieved remarkably enhanced tumor inhibition rate of 56.9% against CT26 colorectal cancer.

Genetically Engineered Cellular Membrane Vesicles as Tailorable Shells for Therapeutics.

Benefiting from the blooming interaction of nanotechnology and biotechnology, biosynthetic cellular membrane vesicles (Bio‐MVs) have shown superior characteristics for therapeutic transportation because of their hydrophilic cavity and hydrophobic bilayer structure, as well as their inherent biocompatibility and negligible immunogenicity. These excellent cell‐like features with specific functional protein expression on the surface can invoke their remarkable ability for Bio‐MVs based recombinant protein therapy to facilitate the advanced synergy in poly‐therapy. To date, various tactics have been developed for Bio‐MVs surface modification with functional proteins through hydrophobic insertion or multivalent electrostatic interactions. While the Bio‐MVs grow through genetically engineering strategies can maintain binding specificity, sort orders, and lead to strict information about artificial proteins in a facile and sustainable way. In this progress report, the most current technology of Bio‐MVs is discussed, with an emphasis on their multi‐functionalities as “tailorable shells” for delivering bio‐functional moieties and therapeutic entities. The most notable success and challenges via genetically engineered tactics to achieve the new generation of Bio‐MVs are highlighted. Besides, future perspectives of Bio‐MVs in novel bio‐nanotherapy are provided.

Isolation and Identification of Pseudo Seven-Coordinate Ru(III) Intermediate Completing the Catalytic Cycle of Ru-bda Type of Water Oxidation Catalysts

Isolation and Identification of Pseudo Seven-Coordinate Ru(III) Intermediate Completing the Catalytic Cycle of Ru-bda Type of Water Oxidation Catalysts

Polymer Vesicles for Antimicrobial Applications.

Polymer vesicles, hollow nanostructures with hydrophilic cavity and hydrophobic membrane, have shown significant potentials in biomedical applications including drug delivery, gene therapy, cancer theranostics, and so forth, due to their unique cell membrane-like structure. Incorporation with antibacterial active components like antimicrobial peptides, etc., polymer vesicles exhibited enhanced antimicrobial activity, extended circulation time, and reduced cell toxicity. Furthermore, antibacterial, and anticancer can be achieved simultaneously, opening a new avenue of the antimicrobial applications of polymer vesicles. This review seeks to highlight the state-of-the-art of antimicrobial polymer vesicles, including the design strategies and potential applications in the field of antibacterial. The structural features of polymer vesicles, preparation methods, and the combination principles with antimicrobial active components, as well as the advantages of antimicrobial polymer vesicles, will be discussed. Then, the diverse applications of antimicrobial polymer vesicles such as wide spectrum antibacterial, anti-biofilm, wound healing, and tissue engineering associated with their structure features are presented. Finally, future perspectives of polymer vesicles in the field of antibacterial is also proposed.

Impact of dimensionality and confinement on reaction dynamics and thermodynamics within 1D and 2D nanostructures.

Confinement has been shown to contribute to the dynamics of small molecules within nanoscale hydrophobic or hydrophilic cavities. Enclosure within a confined space can also influence energy transfer pathways, such as the enhancement of fluorescence over thermal relaxation. In this paper, the effect of confinement on the thermodynamic properties and reaction kinetics of small hydrophobic molecules confined in a soft polymeric template is detailed. A quasi-elastic neutron scattering experiment identified a substantial decrease in translational diffusion of pyrrole after solubilization within a hydrophobic cavity. This decrease in mobility is due to pyrrole's closer packing and increased density under confinement vs the bulk liquid. The decreased mobility and increased density explain the spontaneous polymerization reaction of pyrrole observed within the cavity. The precise characterization of the polymerization kinetics under confinement found that the reaction is independent of pyrrole concentration, consistent with the close packing density. Kinetic data also show that confinement dimensionality finds a thermodynamic expression in the transition state entropy. The dynamics and kinetics experiments reported here offer rare empirical insight into the important influence that cavity geometry places on the reactions they host.

Near-Infrared Photo-controlled Permeability of a Biomimetic Polymersome with Sustained Drug Release and Efficient Tumor Therapy.

Synthetic polymersomes have structure similarity to bio-vesicles and could disassemble in response to stimuli for "on-demand" release of encapsulated cargos. Though widely applied as a drug delivery carrier, the burst release mode with structure complete destruction is usually taken for most responsive polymersomes, which would shorten the effective drug reaction time and impair the therapeutic effect. Inspired by the cell organelles' communication mode via regulating membrane permeability for transportation control, we highlight here a biomimetic polymersome with sustained drug release over a specific period of time via near-infrared (NIR) pre-activation. The polymersome is prepared by the self-assembling amphiphilic diblock copolymer P(OEGMA-co-EoS)-b-PNBOC and encapsulates the hypoxia-activated prodrug AQ4N and upconversion nanoparticle (PEG-UCNP) in its hydrophilic centric cavity. Thirty minutes of NIR pre-activation triggers cross-linking of NBOC and converts the permeability of the polymersome with sustained AQ4N release until 24 h after the NIR pre-activation. The photosensitizer EoS is activated and aggravates environmental hypoxic conditions during a sustained drug release period to boost the AQ4N therapeutic effect. The combination of sustained drug release with concurrent hypoxia intensification results in a highly efficient tumor therapeutic effect both intracellularly and in vivo. This biomimetic polymersome will provide an effective and universal tumor therapeutic approach.

Research Progress on Synthesis and Application of Cyclodextrin Polymers.

Cyclodextrins (CDs) are a series of cyclic oligosaccharides formed by amylose under the action of CD glucosyltransferase that is produced by Bacillus. After being modified by polymerization, substitution and grafting, high molecular weight cyclodextrin polymers (pCDs) containing multiple CD units can be obtained. pCDs retain the internal hydrophobic-external hydrophilic cavity structure characteristic of CDs, while also possessing the stability of polymer. They are a class of functional polymer materials with strong development potential and have been applied in many fields. This review introduces the research progress of pCDs, including the synthesis of pCDs and their applications in analytical separation science, materials science, and biomedicine.

The catalytic machinery of the FAD-dependent AtBBE-like protein 15 for alcohol oxidation: Y193 and Y479 form a catalytic base, Q438 and R292 an alkoxide binding site

Monolignol oxidoreductases are members of the berberine bridge enzyme–like (BBE-like) protein family (pfam 08031) that oxidize monolignols to the corresponding aldehydes. They are FAD-dependent enzymes that exhibit the para-cresolmethylhydroxylase-topology, also known as vanillyl oxidase-topology. Recently, we have reported the structural and biochemical characterization of two monolignol oxidoreductases from Arabidopsis thaliana, AtBBE13 and AtBBE15. Now, we have conducted a comprehensive site directed mutagenesis study for AtBBE15, to expand our understanding of the catalytic mechanism of this enzyme class. Based on the kinetic properties of active site variants and molecular dynamics simulations, we propose a refined, structure-guided reaction mechanism for the family of monolignol oxidoreductases. Here, we propose that this reaction is facilitated stepwise by the deprotonation of the allylic alcohol and a subsequent hydride transfer from the Cα-atom of the alkoxide to the flavin. We describe an excessive hydrogen bond network that enables the catalytic mechanism of the enzyme. Within this network Tyr479 and Tyr193 act concertedly as active catalytic bases to facilitate the proton abstraction. Lys436 is indirectly involved in the deprotonation as this residue determines the position of Tyr193 via a cation-π interaction. The enzyme forms a hydrophilic cavity to accommodate the alkoxide intermediate and to stabilize the transition state from the alkoxide to the aldehyde. By means of molecular dynamics simulations, we have identified two different and distinct binding modes for the substrate in the alcohol and alkoxide state. The alcohol interacts with Tyr193 and Tyr479 while Arg292, Gln438 and Tyr193 form an alkoxide binding site to accommodate this intermediate. The pH-dependency of the activity of the active site variants revealed that the integrity of the alkoxide binding site is also crucial for the fine tuning of the pKa of Tyr193 and Tyr479. Sequence alignments showed that key residues for the mechanism are highly conserved, indicating that our proposed mechanism is not only relevant for AtBBE15 but for the majority of BBE-like proteins.

Компьютерное моделирование и численные исследования пептидных нанотрубок на основе дифенилаланина

The work is devoted to computer studies of the structural and physical properties of such self-organizing structures as peptide nanotubes based on diphenylanine dipeptide with different initial isomers of the left and right chiralities of these dipeptides. The structures under study are considered both with empty anhydrous and with internal cavities filled with water molecules. Molecular models of both ring and helix structures are investigated, which are consistent with the known experimental data. To study the effect of nano-sized clusters of water molecules embedded in the inner hydrophilic cavity on the properties of nanotubes (including the changes in their dipole moments and polarizations), as well as the changes in the structure and properties of water clusters themselves (their dipole moments and polarizations), the surfaces of internal cavities of nanotubes and outer surfaces of water cluster structures for both types of chirality are analyzed. Including using a specially developed method of visual differential analysis of structural features of (bio)macromolecular structures. The obtained results of calculations of a number of physical properties (polarization values, etc.) are given for various cases and analyzed in comparison with the known data. These data are necessary for analyzing the interactions of water molecules with hydrophilic parts of nanotube molecules based on diphenylalanine, such as COO<sup>-</sup> and NH<sub>3</sub><sup>+</sup>, since they determine many properties of the structures under study. A number of the obtained data are planned to be used for further analysis of the possible adhesion and capture of certain medical molecular agents (components) by active layers of diphenylalanine-based nanotubes, which can be useful for creating capsules for targeted delivery of pharmaceuticals and drugs on their basis.

Cyclodextrin-based metal-organic frameworks for pulmonary delivery of curcumin with improved solubility and fine aerodynamic performance.

Pulmonary drug delivery has attracted considerable attention in recent years. However, it is still a major challenge to deliver poorly water-soluble drugs to lungs with good solubility and fine aerodynamic performance. In this study, curcumin was loaded into cyclodextrin-based metal-organic frameworks (CD-MOFs) for pulmonary delivery. Compared with micronized curcumin prepared by jet milling, curcumin-loaded CD-MOFs (Cur-CD-MOFs) exhibited excellent aerodynamic performance, which was attributed to the unique porous structure and lower density of CD-MOFs. The dissolution test showed that the drug release rate of Cur-CD-MOFs was much faster than that of micronized curcumin. The all-atom molecular dynamic simulation showed that curcumin molecules were loaded into the hydrophobic cavities of CD-MOFs or entered into the large hydrophilic cavities to form nanoclusters. The elevated wettability of Cur-CD-MOFs and the unique spatial distribution feature of curcumin in porous interior of CD-MOFs might be favorable for the improved dissolution rate. The DPPH radical scavenging test showed that Cur-CD-MOFs had prominent antioxidant activities. Therefore, CD-MOFs were expected to be promising carriers for pulmonary delivery of poorly water-soluble drugs.

Determination of melamine in milk based on β-cyclodextrin modified carbon nanoparticles via host–guest recognition

Illegal addition of melamine (MEL) to milk has caused serious food safety accident. It is urgent to develop a highly sensitive method for detecting MEL in milk. β-Cyclodextrin with inner hydrophobic and outer hydrophilic cavities have been widely used in smart sensors design. In this study, an “ON-OFF-ON” sensor for MEL detection was constructed based on β-cyclodextrin modified carbon nanoparticles (β-CD-CNPs). The sensor is switched “OFF” when Fe3+ interacts with β-CD-CNPs and switched “ON” when MEL replaces Fe3+. Fluorescence recovery of β-CD-CNPs exhibits good linear correlations with MEL concentration ranging from 10.00 ng/mL ~ 180.00 ng/mL and 180.00 ~ 1000.00 ng/mL, the detection limit is 6.82 ng/mL. The sensor was applied to analysis melamine in milk samples with recovery between 94.80% ~ 102.05%, and RSD bellow 12.61%. The results show that this method can meet the requirements of real sample analysis.

Co-encapsulation of Vitamin C and β-Carotene in liposomes: Storage stability, antioxidant activity, and in vitro gastrointestinal digestion

Vitamin C (VC) and β-Carotene (βC) were selected to produce co-encapsulated liposomes using hydrophilic and hydrophobic cavities simultaneously by ethanol injection method. The results of liposomal structure characterized by particle size, polydispersity index, zeta-potential and transmission electron microscope showed that the microstructure of all liposomal samples was spherical without adhesion or break and the size of VC-βC-loaded liposome (L-VC-βC) was bigger than VC-loaded liposome (L-VC) or βC-loaded liposome (L-βC). The encapsulation efficiency (EE) of VC in L-VC-βC was significantly higher than that in L-VC, and the EE of βC in L-VC-βC had no significant change compared with that in L-βC. The free radical scavenging rate of L-VC-βC was significantly higher than that of L-βC, while it had no significant change compared with that of L-VC. In addition, the storage stability of βC in L-VC-βC improved greatly compared with that in L-βC. Furthermore, the zero order model was applied to understand the release kinetics of βC from L-βC and L-VC-βC in the stomach, whereas the Korsmeyr-Peppas model was chosen to describe the release of βC from two types of liposome in small intestine and their release mechanisms were mainly dominated by Fickian diffusion. It was significant to provide a new idea for using hydrophilic and hydrophobic cavities simultaneously in liposomes to design the multicomponent nutrient delivery system.

Cryo-EM structure of an amyloid fibril formed by full-length human prion protein.

Prion diseases are caused by the misfolding of prion protein (PrP). Misfolded PrP forms protease-resistant aggregates in vivo (PrPSc) that are able to template the conversion of the native form of the protein (PrPC), a property shared by in vitro-produced PrP fibrils. Here we produced amyloid fibrils in vitro from recombinant, full-length human PrPC (residues 23-231) and determined their structure using cryo-EM, building a model for the fibril core comprising residues 170-229. The PrP fibril consists of two protofibrils intertwined in a left-handed helix. Lys194 and Glu196 from opposing subunits form salt bridges, creating a hydrophilic cavity at the interface of the two protofibrils. By comparison with the structure of PrPC, we propose that two α-helices in the C-terminal domain of PrPC are converted into β-strands stabilized by a disulfide bond in the PrP fibril. Our data suggest that different PrP mutations may play distinct roles in modulating the conformational conversion.