Multicomponent Assembly Research Articles

Induction of multicomponent peptide assembly via solvent exchange strategy for enhanced structure and bioactivities

Multicomponent peptides, composed of numerous peptides with various binding sites and functionalities, are increasingly exploited in nanotechnology, biomedicine and functional food applications. This article reported that nucleation and growth of Zein (Z) during solvent exchange could form a confined space to drive spontaneous self-assembly of multicomponent peptides. The self-assembly process began with a concentration gradient generated by the exchange of water and ethanol driving the nucleation of Z to form small aggregates, while multicomponent peptides selectively or anisotropically adsorbed on the Z surface via electrostatic interactions and hydrophobic interactions, participating in and controlling the assembly kinetics, ultimately leading to the formation of well-defined nanoparticles (NPs). The multicomponent peptides-based assemblies could effectively encapsulate the hydrophobic functional agent quercetin (Que), presenting good biocompatibility, antioxidant activity, and preventing DSS-induced colitis. This study elucidated the self-assembly mechanism of multicomponent peptides and demonstrated their synergistic effects with hydrophobic compounds for disease prevention, offering valuable insights into the development of novel functional materials to regulate human health.

Super-Resolution Microscopy as a Versatile Tool in Probing Molecular Assembly.

Molecular assembly is promising in the construction of advanced materials, obtaining structures with specific functions. In-depth investigation of the relationships between the formation, dynamics, structure, and functionality of the specific molecular assemblies is one of the greatest challenges in nanotechnology and chemistry, which is essential in the rational design and development of functional materials for a variety of applications. Super-resolution microscopy (SRM) has been used as a versatile tool for investigating and elucidating the structures of individual molecular assemblies with its nanometric resolution, multicolor ability, and minimal invasiveness, which are also complementary to conventional optical or electronic techniques that provide the direct observation. In this review, we will provide an overview of the representative studies that utilize SRM to probe molecular assemblies, mainly focusing on the imaging of biomolecular assemblies (lipid-based, peptide-based, protein-based, and DNA-based), organic-inorganic hybrid assemblies, and polymer assemblies. This review will provide guidelines for the evaluation of the dynamics of molecular assemblies, assembly and disassembly processes with distinct dynamic behaviors, and multicomponent assembly through the application of these advanced imaging techniques. We believe that this review will inspire new ideas and propel the development of structural analyses of molecular assemblies to promote the exploitation of new-generation functional materials.

Multicomponent Assembling of Aldehydes, N,N‐Dimethylbarbituric Acid, Malononitrile, and Morpholine Into Unsymmetrical Ionic Scaffold and Its Efficient Cyclization

ABSTRACTThe new type of the four‐component tandem Knoevenagel–Michael reaction was discovered: the transformation of arylaldehydes, N,N′‐dimethylbarbituric acid, malononitrile, and morpholine in alcohols and other organic solvents without any other additives at ambient temperature resulted in the selective formation of a new substituted unsymmetrical ionic scaffold with two pharmacology‐active heterocyclic rings. This new four‐component reaction is a simple and efficient route to a previously unknown substituted ionic unsymmetrical scaffold containing N,N′‐dimethylbarbituric acid and morpholine. These ionic scaffolds are promising for various biomedical applications, for example, as anticonvulsants and anti‐inflammatory drugs, as well as anti‐AIDS agents. In this research, we also accomplished the efficient cyclization of morpholin‐4‐ium 5‐(2,2‐dicya‐no‐1‐arylethyl)‐1,3‐dimethyl‐2,6‐di‐oxo‐1,2,3,6‐tetrahydro‐pyrimidin‐4‐olates by the action of NBS–NaOAc system, with the formation of 5,7‐dimethyl‐4,6,8‐trioxo‐2‐aryl‐5,7‐diazaspiro[2.5]octane‐1,1‐dicarbonitriles in 83%–98% yields. In the final stage of our research, direct one‐pot multicomponent transformation of benzaldehydes, N,N′‐dimethylbarbituric acid, and malononitrile into spirobarbituric cyclopropanes was accomplished in 75%–97% yields. The spiro‐barbituric cyclopropanes are potentially active as remedies against various inflammatory, infectious, immunological, or malignant diseases.

Counterion Loss from Charged Surface-Bound Complexes Drives the Formation of Loosely Packed Monolayers.

The functionality of multicomponent self-assembled monolayers (SAMs) can be severely diminished by the segregation of like components into nanoscale domains, a process that maximizes favorable short-range intermolecular interactions. Here, we explore the use of a modular family of sulfur-functionalized metal bis(terpyridine) complexes ([M(tpy-R)2]2+(PF6-)2) to prepare mixed SAMs, considering that the comparable structure, dimensions, and ionic composition of these species should render them interchangeable within the adsorbed surface layer. While surface voltammetry experiments show that these SAMs do exhibit compositions representative of their assembly solutions, they also suggest, in line with previous reports, that adjacent complexes in the monolayer are separated by a gap of ∼ 1 nm. Remarkably, X-ray photoelectron spectroscopy studies reveal no F 1s peak features that would confirm the proliferation of PF6- counterions on the surface. We propose that the loosely packed structure of these SAMs results from the loss or exchange of PF6- counterions, which introduces significant repulsive Coulomb interactions between the adsorbed 2+ charged complexes. The hypothesis is supported by an electrostatic model which indicates that these complexes should form close-packed SAMs if mobile counterions are present. First-principles calculations demonstrate that complex-counterion binding interactions are weakened by charge transfer to the gold substrate, suggesting that this may play an important role in the formation of such low-coverage SAMs. Together, this study raises important questions regarding the assembly, organization, and composition of charged SAMs and highlights new opportunities in the design of multicomponent monolayer assemblies with free volume, for example, to facilitate surface-based reactions or support molecular switches.

Dynamic remodeling of TRPC5 channel–caveolin-1–eNOS protein assembly potentiates the positive feedback interaction between Ca2+ and NO signals

The cell signaling molecules nitric oxide (NO) and Ca2+ regulate diverse biological processes through their closely coordinated activities directed by signaling protein complexes. However, it remains unclear how dynamically the multicomponent protein assemblies behave within the signaling complexes upon the interplay between NO and Ca2+ signals. Here we demonstrate that TRPC5 channels activated by the stimulation of G-protein-coupled ATP receptors mediate Ca2+ influx, that triggers NO production from endothelial NO synthase (eNOS), inducing secondary activation of TRPC5 via cysteine S-nitrosylation and eNOS in vascular endothelial cells. Mutations in the caveolin-1-binding domains of TRPC5 disrupt its association with caveolin-1 and impair Ca2+ influx and NO production, suggesting that caveolin-1 serves primarily as the scaffold for TRPC5 and eNOS to assemble into the signal complex. Interestingly, during ATP receptor activation, eNOS is dissociated from caveolin-1 and in turn directly associates with TRPC5, which accumulates at the plasma membrane dependently on Ca2+ influx and calmodulin. This protein reassembly likely results in a relief of eNOS from the inhibitory action of caveolin-1 and an enhanced TRPC5 S-nitrosylation by eNOS localized in the proximity, thereby facilitating the secondary activation of Ca2+ influx and NO production. In isolated rat aorta, vasodilation induced by acetylcholine was significantly suppressed by the TRPC5 inhibitor AC1903. Thus, our study provides evidence that dynamic remodeling of the protein assemblies among TRPC5, eNOS, caveolin-1, and calmodulin determines the ensemble of Ca2+ mobilization and NO production in vascular endothelial cells.

Nickel-Catalyzed Multicomponent Assembly of Alkynes toward α-CF3-Alkenes.

We disclose an efficient nickel catalytic system for expediting the coupling of alkynes with fluoroalkyl hydrazones and boronic acids, thus facilitating the synthesis of stereospecific α-fluoroalkyl-alkene derivatives. 3H-Pyrazoles might be involved as key intermediates through a nitrogen-releasing process, enabling subsequent coupling with boronic acids to afford 1,2-difunctional alkenes in a highly efficient and step-economical fashion. This tandem platform demonstrates broad functional group tolerance, including complex natural products and drug-like molecules.

Iron Oxide-Doped Carbon Nanoparticles Stabilised with Functionally Modified Hyperbranched Polyglycerol for Cd2+ Sensing and Photodynamic Antibacterial Therapeutic Applications.

Nanoscale materials are being developed from individual particles to multi-component assemblies, with carbon nanomaterials being particularly useful in bioimaging, sensing, and optoelectronics due to their unique optical properties, enhanced by surface passivation and chemical doping. Noble metals are commonly used in conjunction with carbon-based nanomaterials for the synthesis of nanohybrids. Carbon-based materials can function as photosensitizers and effective carriers in photodynamic therapy, enabling the use of combined treatment approaches. The hydrophobicity and agglomeration tendency of carbon nanoparticles pose a drawback. This study is an attempt to overcome these limitations, which involved the synthesis of iron oxide-doped carbon nanoparticles through the carbonisation of citric acid and hexamethylene tetramine, followed by doping them with iron oxide. The as synthesized iron oxide-doped carbon nanoparticles were stabilised with fluorescently modified hyperbranched polyglycerol. The efficacy of these nanoparticles in photodynamic antibacterial therapy and Cd (II) ion sensing was investigated. The selectivity of stabilised nanoparticles against Cd2+ ion is presented in the current study. The current study also compares the antibacterial efficacy of undoped, iron oxide-doped and stabilised nanoparticle systems. The possible toxic effects of the synthesised nanosystems were investigated in order to assess their suitability for biomedical applications and establish their safety profile.

Single-Particle Spectroscopic Chromatography Reveals Heterogeneous RNA Loading and Size Correlations in Lipid Nanoparticles.

Lipid nanoparticles (LNP) have emerged as pivotal delivery vehicles for RNA therapeutics. Previous research and development usually assumed that LNPs are homogeneous in population, loading density, and composition. Such perspectives are difficult to examine due to the lack of suitable tools to characterize these physicochemical properties at the single-nanoparticle level. Here, we report an integrated spectroscopy-chromatography approach as a generalizable strategy to dissect the complexities of multicomponent LNP assembly. Our platform couples cylindrical illumination confocal spectroscopy (CICS) with single-nanoparticle free solution hydrodynamic separation (SN-FSHS) to simultaneously profile population identity, hydrodynamic size, RNA loading levels, and distributions of helper lipid and PEGylated lipid of LNPs at the single-particle level and in a high-throughput manner. Using a benchmark siRNA LNP formulation, we demonstrate the capability of this platform by distinguishing seven distinct LNP populations, quantitatively characterizing size distribution and RNA loading level in wide ranges, and more importantly, resolving composition-size correlations. This SN-FSHS-CICS analysis provides critical insights into a substantial degree of heterogeneity in the packing density of RNA in LNPs and size-dependent loading-size correlations, explained by kinetics-driven assembly mechanisms of RNA LNPs.

Synthesis and Assembly of Photoresponsive Colloidal Tubes.

Inspired by the sophisticated multicomponent and multistage assembly of proteins and their mixtures in living cells, this study rationally designs and fabricates photoresponsive colloidal tubes that can self-assemble and hybrid-assemble when mixed with colloidal spheres and rods. Time-resolved observation and computer simulation reveal that the assembly is driven by phoretic attraction originating from osmotic pressures. These pressures are induced by the chemical concentration gradients generated by the photochemical reaction caused by colloidal tubes in a H2O2 solution under ultraviolet (UV) irradiation. The assembled structure is dictated by the size and shape of the constituent colloids as well as the intensity of the UV irradiation. Additionally, the resulting assembly can undergo self-propelled motion originating from the broken symmetry of the surrounding concentration gradients. This motion can be steered by a magnetic field and used for microscale cargo delivery. The study demonstrates a facile synthesis method for colloidal tubes and highlights their unique potential for controlled, hierarchical self-assembly and hybrid-assembly.

Mechanochemical Assembly of a Nitrile‐Based Directing Group in Arylacetic Acids Using the Passerini 3‐CR: Exploration of the Pd(II)‐Catalyzed meta‐C(sp2)−H Bond Olefination Process

AbstractThe multicomponent assembly of a nitrile‐based directing group in a set of arylacetic acids under solvent‐free mechanochemical conditions using the Passerini 3‐CR and the subsequent Pd(II)‐mediated meta‐C(sp2)−H bond olefination process has been achieved. The protocol demonstrated that removing the DG under mild conditions from the activated Passerini adducts affords a novel N‐(tert‐butyl)‐2‐(2‐cyanophenyl)‐2‐hydroxyacetamide, which can be re‐utilized in the future as DG in other distal activation processes.

Interplay between charge distribution and DNA in shaping HP1 paralog phase separation and localization

The heterochromatin protein 1 (HP1) family is a crucial component of heterochromatin with diverse functions in gene regulation, cell cycle control, and cell differentiation. In humans, there are three paralogs, HP1α, HP1β, and HP1γ, which exhibit remarkable similarities in their domain architecture and sequence properties. Nevertheless, these paralogs display distinct behaviors in liquid-liquid phase separation (LLPS), a process linked to heterochromatin formation. Here, we employ a coarse-grained simulation framework to uncover the sequence features responsible for the observed differences in LLPS. We highlight the significance of the net charge and charge patterning along the sequence in governing paralog LLPS propensities. We also show that both highly conserved folded and less-conserved disordered domains contribute to the observed differences. Furthermore, we explore the potential co-localization of different HP1 paralogs in multicomponent assemblies and the impact of DNA on this process. Importantly, our study reveals that DNA can significantly reshape the stability of a minimal condensate formed by HP1 paralogs due to competitive interactions of HP1α with HP1β and HP1γ versus DNA. In conclusion, our work highlights the physicochemical nature of interactions that govern the distinct phase-separation behaviors of HP1 paralogs and provides a molecular framework for understanding their role in chromatin organization.

Interplay between charge distribution and DNA in shaping HP1 paralog phase separation and localization.

The heterochromatin protein 1 (HP1) family is a crucial component of heterochromatin with diverse functions in gene regulation, cell cycle control, and cell differentiation. In humans, there are three paralogs, HP1α, HP1β, and HP1γ, which exhibit remarkable similarities in their domain architecture and sequence properties. Nevertheless, these paralogs display distinct behaviors in liquid-liquid phase separation (LLPS), a process linked to heterochromatin formation. Here, we employ a coarse-grained simulation framework to uncover the sequence features responsible for the observed differences in LLPS. We highlight the significance of the net charge and charge patterning along the sequence in governing paralog LLPS propensities. We also show that both highly conserved folded and less-conserved disordered domains contribute to the observed differences. Furthermore, we explore the potential co-localization of different HP1 paralogs in multicomponent assemblies and the impact of DNA on this process. Importantly, our study reveals that DNA can significantly reshape the stability of a minimal condensate formed by HP1 paralogs due to competitive interactions of HP1α with HP1β and HP1γ versus DNA. In conclusion, our work highlights the physicochemical nature of interactions that govern the distinct phase-separation behaviors of HP1 paralogs and provides a molecular framework for understanding their role in chromatin organization.

Blueprinting extendable nanomaterials with standardized protein blocks

A wooden house frame consists of many different lumber pieces, but because of the regularity of these building blocks, the structure can be designed using straightforward geometrical principles. The design of multicomponent protein assemblies, in comparison, has been much more complex, largely owing to the irregular shapes of protein structures1. Here we describe extendable linear, curved and angled protein building blocks, as well as inter-block interactions, that conform to specified geometric standards; assemblies designed using these blocks inherit their extendability and regular interaction surfaces, enabling them to be expanded or contracted by varying the number of modules, and reinforced with secondary struts. Using X-ray crystallography and electron microscopy, we validate nanomaterial designs ranging from simple polygonal and circular oligomers that can be concentrically nested, up to large polyhedral nanocages and unbounded straight ‘train track’ assemblies with reconfigurable sizes and geometries that can be readily blueprinted. Because of the complexity of protein structures and sequence–structure relationships, it has not previously been possible to build up large protein assemblies by deliberate placement of protein backbones onto a blank three-dimensional canvas; the simplicity and geometric regularity of our design platform now enables construction of protein nanomaterials according to ‘back of an envelope’ architectural blueprints.

Hybrid functional membranes through layer-by-layer assembly of Ti3C2Tx MXene and gelatin-stabilized calcium phosphate nanospheres

MXene-based membranes exhibit promising properties for various applications; however, pure MXene membranes lack sufficient environmental stability and mechanical strength. This work demonstrates that incorporating gelatin-stabilized amorphous calcium phosphate (Gel-ACP) nanospheres into MXene membranes via layer-by-layer assembly significantly enhances flexibility and mechanical strength while retaining electrical conductivity. A pressure-assisted stacking technique was introduced to efficiently fabricate membranes with tunable thickness not achievable through single filtration steps. The Gel-ACP nanospheres, synthesized by co-precipitation of calcium phosphate within gelatin solution, provided a ductile and cytocompatible reinforcement that augmented the properties of the MXene matrix. Direct mixing of the components led to particle aggregation and non-uniform dispersion. In contrast, controlled layer-by-layer nanostructuring maintained membrane conductivity around 102 Scm−1 while dramatically improving mechanical integrity. The optimized hybrid membranes exhibited specific electromagnetic shielding effectiveness of ∼21,000 dBcm2g−1 and withstood over 90 kPa vacuum pressure without rupture, six times higher than pure MXene membranes. Cytocompatibility was confirmed by the proliferation of human mesenchymal stem cells on the membranes. Moreover, the layered membrane exhibited excellent adhesion to bone-mimicking structures, indicating potential utility for bone tissue engineering. Overall, this work provides new design principles for engineering hybrid membranes through controlled multicomponent assembly to overcome intrinsic limitations and impart multifunctionality.

Non-statistical assembly of multicomponent [Pd2ABCD] cages.

Self-assembled hosts, inspired by biological receptors and catalysts, show application potential in sustainable synthesis, energy conversion and medicine. Implementing multiple functionalities in the form of distinguishable building blocks, however, is difficult without risking narcissistic self-sorting or a statistical mess. Here we report a systematic series of integratively self-assembled heteroleptic cages in which two square-planar PdII cations are bridged by four different bis-pyridyl ligands, A, B, C and D, via synergistic effects to exclusively form a single isomer-the lantern-shaped cage [Pd2ABCD]. This self-sorting goal-forming just one out of 55 possible structures-is reached under full thermodynamic control and can be realized progressively (by combining progenitors, such as [Pd2A2C2] with [Pd2B2D2]), directly from ligands and PdII cations or by mixing all four corresponding homoleptic cages. The rational design of complex multicomponent assemblies that enables the modular incorporation of diverse chemical moieties will advance their applicability in functional nanosystems.

Streamlining the automated discovery of porous organic cages.

Self-assembly through dynamic covalent chemistry (DCC) can yield a range of multi-component organic assemblies. The reversibility and dynamic nature of DCC has made prediction of reaction outcome particularly difficult and thus slows the discovery rate of new organic materials. In addition, traditional experimental processes are time-consuming and often rely on serendipity. Here, we present a streamlined hybrid workflow that combines automated high-throughput experimentation, automated data analysis, and computational modelling, to accelerate the discovery process of one particular subclass of molecular organic materials, porous organic cages. We demonstrate how the design and implementation of this workflow aids in the identification of organic cages with desirable properties. The curation of a precursor library of 55 tri- and di-topic aldehyde and amine precursors enabled the experimental screening of 366 imine condensation reactions experimentally, and 1464 hypothetical organic cage outcomes to be computationally modelled. From the screen, 225 cages were identified experimentally using mass spectrometry, 54 of which were cleanly formed as a single topology as determined by both turbidity measurements and 1H NMR spectroscopy. Integration of these characterisation methods into a fully automated Python pipeline, named cagey, led to over a 350-fold decrease in the time required for data analysis. This work highlights the advantages of combining automated synthesis, characterisation, and analysis, for large-scale data curation towards an accessible data-driven materials discovery approach.

Development of photoluminescent hydrogen-bonded frameworks based on pyromellitic diimide-tethered carboxylic acid hosts and multi-bonding solvent guests

Supramolecular aspects of multi-component hydrogen-bonded assemblies constructed by the combinations of pyromellitic diimide-tethered carboxylic acids and some multiple hydrogen-bonded donor/acceptor solvent molecules are studied.

Dynamic Changes in the Thylakoid Proteome of Cyanobacteria during Light-Regulated Thylakoid Membrane Development

Cyanobacteria were among the oldest organisms to undertake oxygenic photosynthesis and have an essential impact on the atmosphere and carbon/nitrogen cycles on the planet. The thylakoid membrane of cyanobacteria represents an intricate compartment that houses a variety of multi-component (pigment–)protein complexes, assembly factors, and regulators, as well as transporters involved in photosynthetic light reactions, and respiratory electron transport. How these protein components are incorporated into membranes during thylakoid formation and how individual complexes are regulated to construct the functional machinery remains elusive. Here, we carried out an in-depth statistical analysis of the thylakoid proteome data obtained during light-induced thylakoid membrane biogenesis in the model cyanobacterium Synechococcus elongatus PCC 7942. A total of 1581 proteins were experimentally quantified, among which 457 proteins demonstrated statistically significant variations in abundance at distinct thylakoid biogenesis stages. Gene Ontology and KEGG enrichment analysis revealed that predominantly photosystems, light-harvesting antennae, ABC transporters, and pathway enzymes involved in oxidative stress responses and protein folding exhibited notable alternations in abundance between high light and growth light. Moreover, through cluster analysis the 1581 proteins were categorized into six distinct clusters that have significantly different trajectories of the change in their abundance during thylakoid development. Our study provides insights into the physiological regulation for the membrane integration of protein components and functionally linked complexes during the cyanobacterial TM biogenesis process. The findings and analytical methodologies developed in this study may be valuable for studying the global responses of TM biogenesis and photosynthetic acclimation in plants and algae.

Super-assembly strategy based on discretization design for composite lattice metamaterials

The manufacture, multi-component assembly, and connection of composite lattice metamaterials have seriously restricted its large-scale industrial application. For a feasible solution, a super-assembly strategy based on the discretization method has been designed to establish composite lattice metamaterials. They can assemble a wide variety of single- and multi-order super-hybrid structures using a few types of voxel elements. Two kinds of CFRP lattices are successfully fabricated by this strategy. Uniaxial compression responses of the manufactured structures are experimented, and the results have shown good agreement with theoretical predictions and FEA simulations. The deformation and failure modes are analyzed systematically to verify the rationality of this strategy. The assessment of mechanical properties has demonstrated that the proposed design strategy is quite superior to existing other lattices prepared using analogical discrete-assembly techniques. This work has shown promising potential for industrial production with the advantages of low cost, high flexibility, high efficiency, standardization, and large-scale.

Synthesis and characterization of new biocatalyst based on LDH functionalized with l-asparagine amino acid for the synthesis of tri-substituted derivatives of 2, 4, 5-(H1)-imidazoles

In this study, a new and recyclable biocatalyst (MgAl CO3-LDH@Asn) was synthesized by immobilizing l-asparagine amino acid (Asn) on the surface of 3-(chloropropyl)-trimethoxysilane modified MgAl CO3-layered double hydroxide (LDH). The physicochemical properties of the samples were identified by Fourier transform infrared (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray (EDX), and thermogravimetric analysis (TGA) techniques. The MgAl CO3-LDH@Asn was employed in the multi-component assembly process for the synthesis of tri-substituted derivatives of 2,4,5-(H1)-imidazoles from benzyl, various benzaldehyde derivatives, and ammonium acetate. For optimizing the reaction, the main factors, including the amount of MgAl CO3-LDH@Asn, type of solvent, reaction time, and temperature were evaluated. The optimum conditions of the model reaction were achieved using 20 mg of MgAl CO3-LDH@Asn biocatalyst in ethanol solvent after 20 min at reflux temperature. According to the findings above, the results indicated that high-yield products are achieved within a short time frame. Moreover, the high catalytic activity of the MgAl CO3-LDH@Asn was maintained for four cycles without significantly diminishing its performance.