Spatiotemporal organization of coacervate microdroplets

Liquid–liquid phase separation (LLPS) has emerged as a new paradigm in the fields of soft matter, colloid chemistry, prebiotic chemistry, and cell biology. As phase separation is a dynamic assembly process, how to spatiotemporally regulate the assembly and disassembly of these micrometre-sized droplets, which are referred as biomolecular condensates in biology is essential for their diverse applications in various disciplines. Herein, we discuss recent advances in the spatiotemporal control of phase separation using different physical tools and external environmental stimuli in bulk solutions and living cells. Specifically, the exploration of phase transition in a compartmentalized protocellular system, which can bridge the gap between synthetic and intracellular LLPS systems, is summarized, and the challenges and future research directions are discussed.     

Heterochromatin Protein HP1α Gelation Dynamics Revealed by Solid‐State NMR Spectroscopy

Heterochromatin protein 1α (HP1α) undergoes liquid–liquid phase separation (LLPS) and forms liquid droplets and gels in vitro, properties that also appear to be central to its biological function in heterochromatin compaction and regulation. Here we use solid‐state NMR spectroscopy to track the conformational dynamics of phosphorylated HP1α during its transformation from the liquid to the gel state. Using experiments designed to probe distinct dynamic modes, we identify regions with varying mobilities within HP1α molecules and show that specific serine residues uniquely contribute to gel formation. The addition of chromatin disturbs the gelation process while preserving the conformational dynamics within individual bulk HP1α molecules. Our study provides a glimpse into the dynamic architecture of dense HP1α phases and showcases the potential of solid‐state NMR to detect an elusive biophysical regime of phase separating biomolecules.     

Preparation of polypropylene microspheres for selective laser sintering via thermal‐induced phase separation: Roles of liquid–liquid phase separation and crystallization

Although selective laser sintering (SLS) has been widely applied in many fields, more research work is needed to develop proper polymer microspheres for SLS. Thermal‐induced phase separation (TIPS) is a facile way but rarely reported to prepare the polymer microspheres. The roles of liquid–liquid phase separation (LLPS) and crystallization in the TIPS process are not clear. In this study, proper polypropylene (PP) microspheres for SLS are successfully prepared via TIPS with xylene. The diameters and morphologies of these PP microspheres can be regulated easily by changing the PP concentration and the quench temperature. The large undercooling drives the solution into the metastable LLPS region and produces PP microspheres with smooth surfaces. The PP crystallization occurs both on the LLPS interface and inside the polymer‐rich phase when the solution is quenched to a temperature near the binodal line, and the tiny bent lamellae are formed on the microsphere surface. At higher temperature only PP crystallization occurs, which results in the formation of PP particles consisting of packed lamellae. The PP microspheres prepared here are suitable for SLS and promote the development of SLS potentially. © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2017 , 55, 320–329     

Liquid-liquid phase transition of protein aqueous solutions isothermally induced by protein cross-linking

We experimentally demonstrated that liquid-liquid phase separation (LLPS) of protein aqueous solutions can be induced by isothermal protein oligomerization. This phenomenon is analogous to LLPS induced by the polymerization of small organic molecules in solution. Specifically, using glutaraldehyde for protein cross-linking, we observed the formation of protein-rich liquid droplets for bovine serum albumin and chicken egg lysozyme at 25 degrees C. These droplets evolved into cross-linked protein microspheres. If the aqueous solutions of the protein monomer do not show LLPS at temperatures lower than the oligomerization temperature, protein-rich droplets are not observed. We experimentally linked the formation of these droplets to the increase of LLPS temperature during protein oligomerization. When macroscopic aggregation competes with LLPS, a rationale choice of pH, polyethylene glycol, and salt concentrations can be used to favor LLPS relative to aggregation. Although glutaraldehyde has been extensively used to cross-link protein molecules, to our knowledge, its use in homogeneous aqueous solutions to induce LLPS has not been previously described. This work contributes to the fundamental understanding of both phase transitions of protein solutions and the morphology of protein condensed phases. It also provides guidance for the development of new methods based on mild experimental conditions for the preparation of protein-based materials.     

Supramolecular colloidal motors via chemical self-assembly

A colloidal motor can convert energy stored in the environment to achieve mechanical motion and exhibit dynamic behaviors in fluids. To overcome the challenges presented to a colloidal motor, controlled molecular self-assembly technology provides new opportunities for the precise fabrication of various nanoarchitectures and facilitates fundamental research on rational design, multifunctionalization, propulsion, and controlled movement of colloidal motors. These molecular assembled colloidal motors, also called supramolecular colloidal motors, can perform special tasks at the micro- and nanoscale in the fields of biomedicine, nanotechnology, and environmental remediation. In this feature article, we first introduce the recent progress of controllable self-assembly of spatially asymmetric supramolecular colloidal motors with variable sizes, structures, and functions and discuss the relationship between structure and propulsion. Next, we review the research progress of this type of colloidal motors in biomedical and environmental fields. Finally, we propose the challenges of the supramolecular colloidal motors and future development direction.     

Porphyrin/Ionic‐Liquid Co‐assembly Polymorphism Controlled by Liquid–Liquid Phase Separation

Understanding and controlling multicomponent co‐assembly is of primary importance in different fields, such as materials fabrication, pharmaceutical polymorphism, and supramolecular polymerization, but these aspects have been a long‐standing challenge. Herein, we discover that liquid–liquid phase separation (LLPS) into ion‐cluster‐rich and ion‐cluster‐poor liquid phases is the first step prior to co‐assembly nucleation based on a model system of water‐soluble porphyrin and ionic liquids. The LLPS‐formed droplets serve as the nucleation precursors, which determine the resulting structures and properties of co‐assemblies. Co‐assembly polymorphism and tunable supramolecular phase transition behaviors can be achieved by regulating the intermolecular interactions at the LLPS stage. These findings elucidate the key role of LLPS in multicomponent co‐assembly evolution and enable it to be an effective strategy to control co‐assembly polymorphism as well as supramolecular phase transitions.     

“可控自组装体系及其功能化”重大研究计划取得系列重要研究成果

In 2005, the Science magazine, in commemorating the 125th anniversary of its founding, proposed 25 most challenging scientific issues in the future. One of the issues, "How far can we push chemical self-assembly?" has attracted the attention of scientists all over the world. During the 11th Five Year Plan period, NSFC convened scientists from various fields and proposed a major research project, "Controllable self-assembly system and its functionalization". Since the implementation of this project, Chinese scientists have developed and recognized a variety of noncovalent interactions, constructed numerous assembly building blocks with the "Chinese label", established a new assembly method similar to an organic "name reaction", realized the functions of multi-component and multi-level assemblies, and constructed a batch of controllable self-assembly systems having scientific importance and potential practical value. They have achieved great leap forward from following to original innovation, and brought the research of chemical self-assembly in China to move forward to the center of international stage. In this study, we examined the overall scientific goals, general layout of the plan, and ideas for implementation of the major research program "Controllable Self-Assembly System and its Functionalization", as well as an array of significant research accomplishments made possible through the funding received by the program.     

Porphyrin/Ionic-Liquid Co-assembly Polymorphism Controlled by Liquid–Liquid Phase Separation

Understanding and controlling multicomponent co-assembly is of primary importance in different fields, such as materials fabrication, pharmaceutical polymorphism, and supramolecular polymerization, but these aspects have been a long-standing challenge. Herein, we discover that liquid–liquid phase separation (LLPS) into ion-cluster-rich and ion-cluster-poor liquid phases is the first step prior to co-assembly nucleation based on a model system of water-soluble porphyrin and ionic liquids. The LLPS-formed droplets serve as the nucleation precursors, which determine the resulting structures and properties of co-assemblies. Co-assembly polymorphism and tunable supramolecular phase transition behaviors can be achieved by regulating the intermolecular interactions at the LLPS stage. These findings elucidate the key role of LLPS in multicomponent co-assembly evolution and enable it to be an effective strategy to control co-assembly polymorphism as well as supramolecular phase transitions.     

Active structuring of colloids through field-driven self-assembly

In recent years self-assembly has become progressively more “active”, i.e. the focus of research gradually has shifted towards field-manipulation of matter in order to form temporary states rather than creating static architectures. The desire for time-programmed control of materials certainly originates from the unmatched complexity of natural systems that orchestrate multiple components across length scales. Although artificial self-assembly still lacks control comparable to natural systems, there has been impressive progress in a concerted approach from physicists, chemists, biologists, and engineers. This review summarizes the current trend in colloidal assembly advancing from static assembly of isotropic particles towards active structuring of anisotropic particles with heterogeneous (patchy) surfaces, and ultimately, to complex behavior in dissipative dynamic systems. We focus both on the formation of static structures and on temporary states due to response to magnetic, electric, or optic stimulation. We give examples of nano- and microparticle assembly where the temporary state may adopt equilibrium order or a continuously changing dynamic pattern.     

制备方法对聚合物共混物相分离与黏弹弛豫的影响

采用小角激光光散射(SALLS)和动态流变方法研究了通过不同制备方法得到的等规聚丙烯/乙丙橡胶共混物(iPP/EPR)的相分离行为与黏弹行为.依据Cahn-Hilliard-Cook理论分析了熔融共混和溶液共混法制备的质量比为60/40和40/60的iPP/EPR共混物在恒温相分离早期的动力学,发现熔融共混iPP/EPR具有更大的表观扩散系数(Dapp).相分离中后期的实验结果表明,当相区尺寸增长程度相同时,熔融共混试样所用时间更短.表明熔融共混iPP/EPR试样具有更快的相分离速率.动态流变测试结果表明,与溶液共混相比,熔融共混试样具有更快的松弛速率.考虑到相分离过程实质是由高分子链的运动与扩散所控制,两种方法制备的iPP/EPR共混物相分离速率的差异应归于其分子链运动能力的不同.     

Structural Insight into IAPP-Derived Amyloid Inhibitors and Their Mechanism of Action

Designed peptides derived from the islet amyloid polypeptide (IAPP) cross-amyloid interaction surface with Aβ (termed interaction surface mimics or ISMs) have been shown to be highly potent inhibitors of Aβ amyloid self-assembly. However, the molecular mechanism of their function is not well understood. Using solution-state and solid-state NMR spectroscopy in combination with ensemble-averaged dynamics simulations and other biophysical methods including TEM, fluorescence spectroscopy and microscopy, and DLS, we characterize ISM structural preferences and interactions. We find that the ISM peptide R3-GI is highly dynamic, can adopt a β-like structure, and oligomerizes into colloid-like assemblies in a process that is reminiscent of liquid–liquid phase separation (LLPS). Our results suggest that such assemblies yield multivalent surfaces for interactions with Aβ40. Sequestration of substrates into these colloid-like structures provides a mechanistic basis for ISM function and the design of novel potent anti-amyloid molecules.     

基于有机纳米纤维薄膜的荧光化学传感

薄膜荧光化学传感提供了一种固相、便携、易操作的气相分子检测技术,在环境、安全、生物医学、健康监测等领域具有重要的应用价值和发展前景.基于本课题组在超分子自组装构建n-型有机半导体苝二酰亚胺衍生物(PTCDI)一维纳米纤维及其荧光薄膜检测胺类等气相分子领域研究,结合其他课题组工作,本文阐述了该类纳米纤维多孔薄膜在结构调控,荧光传感应用性能、机制和意义方面的研究进展.同时,也介绍了本课题组在p型有机半导体咔唑角亚乙炔四环(ACTC)和咔唑三聚体等在本领域的进展,最后对未来挑战和发展方向进行了展望.     

Pressure Sensitivity of SynGAP/PSD-95 Condensates as a Model for Postsynaptic Densities and Its Biophysical and Neurological Ramifications

Biomolecular condensates consisting of proteins and nucleic acids can serve critical biological functions, so that some condensates are referred as membraneless organelles. They can also be disease-causing, if their assembly is misregulated. A major physicochemical basis of the formation of biomolecular condensates is liquid–liquid phase separation (LLPS). In general, LLPS depends on environmental variables, such as temperature and hydrostatic pressure. The effects of pressure on the LLPS of a binary SynGAP/PSD-95 protein system mimicking postsynaptic densities, which are protein assemblies underneath the plasma membrane of excitatory synapses, were investigated. Quite unexpectedly, the model system LLPS is much more sensitive to pressure than the folded states of typical globular proteins. Phase-separated droplets of SynGAP/PSD-95 were found to dissolve into a homogeneous solution already at ten-to-hundred bar levels. The pressure sensitivity of SynGAP/PSD-95 is seen here as a consequence of both pressure-dependent multivalent interaction strength and void volume effects. Considering that organisms in the deep sea are under pressures up to about 1 kbar, this implies that deep-sea organisms have to devise means to counteract this high pressure sensitivity of biomolecular condensates to avoid harm. Intriguingly, these findings may shed light on the biophysical underpinning of pressure-related neurological disorders in terrestrial vertebrates.     

Fluctuation‐assisted crystallization of an iPP/PEOc polymer alloy prepared on a single multicatalyst reactor granule

The new fluctuation‐assisted mechanism for nucleation and crystallization in the isotactic polypropylene/poly(ethylene‐co‐octene) alloy has been studied. We found that the liquid–liquid phase separation (LLPS) had a dominant influence on the crystallization kinetics through the nucleation process. After LLPS, the nucleation of crystallization mainly occurred at the interface of the phase‐separated domains. It is because that the concentration fluctuations of the LLPS induced the motion of polymer chains and possibly some segmental alignment and/or orientation in the concentration gradient regions through interdiffusion, which could assist the formation of nuclei for crystallization. In other words, the usual nucleation energy barrier could be overcome (or at least partially) by the concentration fluctuation growth of LLPS in the unstable regions. This could be viewed as a new kind of heterogeneous nucleation and could be an addition to the regular nucleation and growth mechanism for crystallization. © 2008 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 47: 166–172, 2009     

Multilayer onion-like vesicles self-assembled from amphiphilic hyperbranched multiarm copolymers via simulation

Multilayer onion-like vesicles are valuable in cell mimics and biomedicine fields. However, as an excellent self-assembly precursor, the formation of multilayer onion-like vesicles by the self-assembly of hyperbranched multiarm copolymers (HMCs) were not reported due to the complex self-assembly dynamics. In this article, the self-assembly behavior of multilayer onion-like vesicles from HMCs was systematically investigated using dissipative particle dynamics simulation. The formation conditions for different kinds of vesicles were disclosed through the construction of the morphological phase diagram. Moreover, the formation mechanisms of the onion-like vesicles with different layers were revealed. We observed that it is the fusion mechanism in low concentrations and the molecular rearrangement mechanism in high concentrations. For low concentration, the law between the number of the membrane layers and the morphology of the aggregates in the fusion process was disclosed. Meanwhile, the membrane of the onion-like vesicles self-assembled from HMCs is monolayer structure and the thickness of each layer is decreased in sequence from inside to outside. The current observations have important guiding significance for its application in drug delivery systems.     

Remodeling of the Fibrillation Pathway of α-Synuclein by Interaction with Antimicrobial Peptide LL-III

Liquid-liquid phase separation (LLPS) has emerged as a key mechanism for intracellular organization, and many recent studies have provided important insights into the role of LLPS in cell biology. There is also evidence that LLPS is associated with a variety of medical conditions, including neurodegenerative disorders. Pathological aggregation of α-synuclein, which is causally linked to Parkinson's disease, can proceed via droplet condensation, which then gradually transitions to the amyloid state. We show that the antimicrobial peptide LL-III is able to interact with both monomers and condensates of α-synuclein, leading to stabilization of the droplet and preventing conversion to the fibrillar state. The anti-aggregation activity of LL-III was also confirmed in a cellular model. We anticipate that studying the interaction of antimicrobial-type peptides with liquid condensates such as α-synuclein will contribute to the understanding of disease mechanisms (that arise in such condensates) and may also open up exciting new avenues for intervention.     

Oil-water interfacial self-assembly: a novel strategy for nanofilm and nanodevice fabrication

How to integrate individual nanostructures into macroscopic thin films has become one of the most intriguing fields in nanoscience and nanotechnology due to the unique properties and important applications of these functional films. Since being discovered in 2004, oil-water interfacial self-assembly of nanostructures has become a novel strategy for fabrication of nanofilms. It is a powerful bottom-up approach for film fabrication due to the low cost and high efficiency, and is simple and universal for almost all low-dimensional nanostructures. In this article, we provide a critical review of the state-of-the-art research activities in this burgeoning self-assembly strategy. We first discuss the thermodynamic mechanism of the oil-water interfacial self-assembly, then the self-assembly of various low-dimensional nanostructures including nanoparticles, one-dimensional (1D) nanostructures, two-dimensional (2D) nanostructures at an oil-water interface developed so far to fabricate high-quality nanofilms. Finally, we present some progress on the construction of functional nanofilm-based nanodevices from this novel strategy based on our research. We conclude this review with critical comments on advantages and the experimental challenges, and further propose the future research and development of this self-assembly strategy for nanodevice construction (105 references).     

Complex collective dynamics of active torque-driven colloids at interfaces

Modern self-assembly techniques aiming to produce complex structural order or functional diversity often rely on non-equilibrium conditions in the system. Light, electric, or magnetic fields are predominantly used to modify interaction profiles of colloidal particles during self-assembly or induce complex out-of-equilibrium dynamic ordering. The energy injection rate, properties of the environment are important control parameters that influence the outcome of active (dynamic) self-assembly. The current review is focused on a case of collective dynamics and self-assembly of particles with externally driven torques coupled to a liquid or solid interface. The complexity of interactions in such systems is further enriched by strong hydrodynamic coupling between particles. Unconventionally ordered dynamic self-assembled patterns, spontaneous symmetry breaking phenomena, self-propulsion, and collective transport have been reported in torque-driven colloids. Some of the features of the complex collective behavior and dynamic pattern formation in those active systems have been successfully captured in simulations.     

Caught in Action: Visualizing Dynamic Nanostructures Within Supramolecular Systems Chemistry

Supramolecular systems chemistry has been an area of active research to develop nanomaterials with life-like functions. Progress in systems chemistry relies on our ability to probe the nanostructure formation in solution. Often visualizing the dynamics of nanostructures which transform over time is a formidable challenge. This necessitates a paradigm shift from dry sample imaging towards solution-based techniques. We review the application of state-of-the-art techniques for real-time, in situ visualization of dynamic self-assembly processes. We present how solution-based techniques namely optical super-resolution microscopy, solution-state atomic force microscopy, liquid-phase transmission electron microscopy, molecular dynamics simulations and other emerging techniques are revolutionizing our understanding of active and adaptive nanomaterials with life-like functions. This Review provides the visualization toolbox and futuristic vision to tap the potential of dynamic nanomaterials.     

Observation of liquid–liquid phase separation of ataxin-3 and quantitative evaluation of its concentration in a single droplet using Raman microscopy

Liquid–liquid phase separation (LLPS) plays an important role in a variety of biological processes and is also associated with protein aggregation in neurodegenerative diseases. Quantification of LLPS is necessary to elucidate the mechanism of LLPS and the subsequent aggregation process. In this study, we showed that ataxin-3, which is associated with Machado–Joseph disease, exhibits LLPS in an intracellular crowding environment mimicked by biopolymers, and proposed that a single droplet formed in LLPS can be quantified using Raman microscopy in a label-free manner. We succeeded in evaluating the protein concentration and identifying the components present inside and outside a droplet using the O–H stretching band of water as an internal intensity standard. Only water and protein were detected to be present inside droplets with crowding agents remaining outside. The protein concentration in a droplet was dependent on the crowding environment, indicating that the protein concentration and intracellular environment should be considered when investigating LLPS. Raman microscopy has the potential to become a powerful technique for clarifying the chemical nature of LLPS and its relationship with protein aggregation.

Liquid–liquid phase separation (LLPS) plays an important role in a variety of biological processes. We have established a method to quantify a single droplet formed by LLPS using the Raman band of water as an internal standard.