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

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

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

Nanoparticulate Nonviral Agent for the Effective Delivery of pDNA and siRNA to Differentiated Cells and Primary Human T Lymphocytes

Delivery of polynucleotides such as plasmid DNA (pDNA) and siRNA to nondividing and primary cells by nonviral vectors presents a considerable challenge. In this contribution, we introduce a novel type of PDMAEMA-based star-shaped nanoparticles that (i) are efficient transfection agents in clinically relevant and difficult-to-transfect human cells (Jurkat T cells, primary T lymphocytes) and (ii) can efficiently deliver siRNA to human primary T lymphocytes resulting to more than 40% silencing of the targeted gene. Transfection efficiencies achieved by the new vectors in serum-free medium are generally high and only slightly reduced in the presence of serum, while cytotoxicity and cell membrane disruptive potential at physiological pH are low. Therefore, these novel agents are expected to be promising carriers for nonviral gene transfer. Moreover, we propose a general design principle for the construction of polycationic nanoparticles capable of delivering nucleic acids to the above-mentioned cells

Nondestructive Light-Initiated Tuning of Layer-by-Layer Microcapsule Permeability

A nondestructive way to achieve remote, reversible, light-controlled tunable permeability of ultrathin shell microcapsules is demonstrated in this study. Microcapsules based on poly{[2-(methacryloyloxy)ethyl] trimethylammonium iodide} (PMETAI) star polyelectrolyte and poly(sodium 4-styrenesulfonate) (PSS) were prepared by a layer-by-layer (LbL) technique. We demonstrated stable microcapsules with controlled permeability with the arm number of a star polymer having significant effect on the assembly structure: the PMETAI star with 18 arms shows a more uniform and compact assembly structure. We observed that in contrast to regular microcapsules from linear polymers, the permeability of the star polymer microcapsules could be dramatically altered by photoinduced transformation of the trivalent hexacyanocobaltate ions into a mixture of mono- and divalent ions by using UV irradiation. The reversible contraction of PMETAI star polyelectrolyte arms and the compaction of star polyelectrolytes in the presence of multivalent counterions are considered to cause the dramatic photoinduced changes in microcapsule properties observed here. Remarkably, unlike the current mostly destructive approaches, the light-induced changes in microcapsule permeability are completely reversible and can be used for light-mediated loading/unloading control of microcapsules

Multiresponsive Microcapsules Based on Multilayer Assembly of Star Polyelectrolytes

Star polyelectrolytes (poly­(<i>N</i>,<i>N</i>-dimethylaminoethyl methacrylate) (PDMAEMA)) with dual (temperature and pH) responsive properties were utilized to fabricate multiresponsive microcapsules via layer-by-layer (LbL) assembly. The LbL microcapsules are very robust and uniform, with higher stability and different internal structure compared with conventional microcapsules based on linear polyelectrolytes. Ionic strength in the polyelectrolyte solution during the microcapsule assembly process has a significant influence on the thickness and permeability of microcapsules. With increasing pH, the permeability of microcapsules decreases, and the transition from “open” to “closed” state for target molecules can be achieved within a narrow pH range (from pH 7 to 8). On the other hand, the overall size and permeability of the microcapsules decrease with increasing temperature (with a shrinkage of 54% in diameter at 60 °C compared with room temperature), thus allowing to reversibly load and unload the microcapsules with high efficiency. The organization and interaction of star polyelectrolytes within confined multilayer structure are the main driving forces for the responsiveness to external stimuli. The multiresponsive LbL microcapsules represent a novel category of smart microstructures as compared to traditional LbL microcapsules with “one-dimensional” response to a single stimulus, and they also have the potential to mimic the complex responsive microstructures found in nature and find applications in drug delivery, smart coatings, microreactors, and biosensors

Thermo-Induced Limited Aggregation of Responsive Star Polyelectrolytes

Poly­(<i>N</i>,<i>N</i>-dimethylaminoethyl methacrylate) (PDMAEMA) star polyelectrolytes with dual thermo- and pH-responsive properties have been studied by <i>in situ</i> small-angle neutron scattering at different temperatures and pH conditions in order to reveal their conformational changes in semidilute solution. At pH values close to the p<i>K</i>_a, all PDMAEMA stars studied here are partially charged and show a core–shell quasi-micellar morphology caused by microphase separation with a collapsed core region with high monomer density and a hydrated loosely packed shell region. Upon increasing the temperature, the PDMAEMA star polyelectrolytes first experience a contraction in the shell region while the core size remains almost unchanged, and then start to form limited intermolecular aggregates. With decreasing pH values, the transition temperature increases and the size of the aggregates decreases (average aggregation number changes from 10 to 3). We suggest that these changes are triggered by the decrease in solvent quality with increasing temperature, which leads to the transition from an electrostatically dominated regime to a regime dominated by hydrophobic interactions. The observed phenomenon is in striking contrast to the phase behavior of linear PDMAEMA polyelectrolytes, which show macrophase separation with increasing temperature under the same conditions

Hidden Structural Features of Multicompartment Micelles Revealed by Cryogenic Transmission Electron Tomography

The demand for ever more complex nanostructures in materials and soft matter nanoscience also requires sophisticated characterization tools for reliable visualization and interpretation of internal morphological features. Here, we address both aspects and present synthetic concepts for the compartmentalization of nanoparticle peripheries as well as their <i>in situ</i> tomographic characterization. We first form negatively charged spherical multicompartment micelles from ampholytic triblock terpolymers in aqueous media, followed by interpolyelectrolyte complex (IPEC) formation of the anionic corona with bis-hydrophilic cationic/neutral diblock copolymers. At a 1:1 stoichiometric ratio of anionic and cationic charges, the so-formed IPECs are charge neutral and thus phase separate from solution (water). The high chain density of the ionic grafts provides steric stabilization through the neutral PEO corona of the grafted diblock copolymer and suppresses collapse of the IPEC; instead, the dense grafting results in defined nanodomains oriented perpendicular to the micellar core. We analyze the 3D arrangements of the complex and purely organic compartments, <i>in situ</i>, by means of cryogenic transmission electron microscopy (cryo-TEM) and tomography (cryo-ET). We study the effect of block lengths of the cationic and nonionic block on IPEC morphology, and while 2D cryo-TEM projections suggest similar morphologies, cryo-ET and computational 3D reconstruction reveal otherwise hidden structural features, <i>e.g.</i>, planar IPEC brushes emanating from the micellar core

Multicompartment Micelles with Adjustable Poly(ethylene glycol) Shell for Efficient <i>in Vivo</i> Photodynamic Therapy

We describe the preparation of well-defined multicompartment micelles from polybutadiene-<i>block</i>-poly(1-methyl-2-vinyl pyridinium methyl sulfate)-<i>block</i>-poly(methacrylic acid) (BVqMAA) triblock terpolymers and their use as advanced drug delivery systems for photodynamic therapy (PDT). A porphyrazine derivative was incorporated into the hydrophobic core during self-assembly and served as a model drug and fluorescent probe at the same time. The initial micellar corona is formed by negatively charged PMAA and could be gradually changed to poly(ethylene glycol) (PEG) in a controlled fashion through interpolyelectrolyte complex formation of PMAA with positively charged poly(ethylene glycol)-<i>block</i>-poly(l-lysine) (PLL-<i>b</i>-PEG) diblock copolymers. At high degrees of PEGylation, a compartmentalized micellar corona was observed, with a stable bottlebrush-on-sphere morphology as demonstrated by cryo-TEM measurements. By <i>in vitro</i> cellular experiments, we confirmed that the porphyrazine-loaded micelles were PDT-active against A549 cells. The corona composition strongly influenced their <i>in vitro</i> PDT activity, which decreased with increasing PEGylation, correlating with the cellular uptake of the micelles. Also, a PEGylation-dependent influence on the <i>in vivo</i> blood circulation and tumor accumulation was found. Fully PEGylated micelles were detected for up to 24 h in the bloodstream and accumulated in solid subcutaneous A549 tumors, while non- or only partially PEGylated micelles were rapidly cleared and did not accumulate in tumor tissue. Efficient tumor growth suppression was shown for fully PEGylated micelles up to 20 days, demonstrating PDT efficacy <i>in vivo</i>