Submitted / under review
Giacomo Bartolucci, Adriana Calaça Serrão, Philipp Schwintek, Alexandra Kühnlein, Yash Rana, Philipp Janto, Dorothea Hofer, Christof B. Mast, Dieter Braun, and Christoph A. Weber
The emergence of functional oligonucleotides on early Earth required a molecular selection mechanism to screen for specific sequences with prebiotic functions. Cyclic processes such as daily temperature oscillations were ubiquitous in this environment and could trigger oligonucleotide phase separation. Here, we propose sequence selection based on phase separation cycles realized through sedimentation in a system subjected to the feeding of oligonucleotides. Using theory and experiments with DNA, we show sequence-specific enrichment in the sedimented dense phase, in particular of short 22-mer DNA sequences. The underlying mechanism selects for complementarity, as it enriches sequences that tightly interact in the condensed phase through base-pairing. Our mechanism also enables initially weakly biased pools to enhance their sequence bias or to replace the most abundant sequences as the cycles progress. Our findings provide an example of a selection mechanism that may have eased screening for the first auto-catalytic self-replicating oligonucleotides.
Alexander M. Bergmann, Jonathan Bauermann, Giacomo Bartolucci, Carsten Donau, Michele Stasi, Anna-Lena Holtmannspötter, Frank Jülicher, Christoph A. Weber, Job Boekhoven
Liquid-liquid phase separation is the process in which two immiscible liquids demix. This spontaneous phenomenon yields spherical droplets that eventually coarsen to one large, stable droplet governed by the principle of minimal free energy. In chemically fueled phase separation, the formation of phase-separating molecules is coupled to a fuel-driven, nonequilibrium reaction cycle. Chemically fueled phase separation yields dissipative structures sustained by a continuous fuel conversion. Such dissipative structures are ubiquitous in biology but poorly understood as they are governed by non-equilibrium thermodynamics. Here, we bridge the gap between passive, close-to-equilibrium, and active, dissipative structures with chemically fueled phase separation. We observe that spherical, active droplets can transition into a new morphology—a liquid, spherical shell of droplet material. A spherical shell would be highly unstable at equilibrium. Only by continuously converting chemical energy, this dissipative structure can be sustained. We demonstrate the transition mechanism, which is related to the activation of a product outside of the droplet, and the deactivation within the droplets leading to gradients of droplet material. We characterize how far out of equilibrium the spherical shell state is and the chemical power necessary to sustain it. Our work suggests new avenues for assembling complex stable morphologies, which might already be exploited to form membraneless organelles by cells.
Giacomo Bartolucci, Thomas C.T. Michaels, Christoph A. Weber
Interactions among proteins in living cells can lead to molecular assemblies of different sizes and large-scale coexisting phases formed via phase separation. Both are essential for the spatial organization of cells and for regulating biological function and dysfunction. A key challenge is understanding the interplay between molecular assembly and phase separation. However, a corresponding theoretical framework that relies on thermodynamic principles is lacking. Here, we present a non-equilibrium thermodynamic theory for a multi-component mixture that contains assemblies of different sizes, which can form, dissolve, and phase-separate from the solvent. We show that the size distributions of assemblies differ between the phases and that the dense phase can gelate. Moreover, we unravel the mechanisms involved in growth and compositional changes of the coexisting phases during assembly kinetics. Our theory can explain how molecular assembly is intertwined with phase separation, and our results are consistent with recent experimental observations on protein phase separation.
Ivar Svalheim Haugerud, Pranay Jaiswal, Christoph A. Weber
Recent experimental studies suggest that wet-dry cycles and coexisting phases can each strongly alter chemical processes. The mechanisms of why and to which degree chemical processes are altered when subject to evaporation and condensation are unclear. To close this gap, we developed a theoretical framework for non-dilute chemical reactions subject to non-equilibrium conditions of evaporation or condensation. We find that evaporation and condensation can each accelerate the initial rate of chemical processes by more than an order of magnitude, depending on the substrate-solvent interaction. When maintaining the system out of equilibrium by wet-dry cycles, the cycle frequency controls the chemical turnover. Strikingly, there is resonance behavior in the cycle frequency where the turnover is maximal. This resonance behavior enables wet-dry cycles to select specific chemical reactions suggesting a potential mechanism for chemical evolution in prebiotic soups at early Earth.
Jonathan Bauermann, Giacomo Bartolucci, Job Boekhoven, Christoph A. Weber, Frank Jülicher
We study a chemically active binary mixture undergoing phase separation and show that under non-equilibrium conditions, stable liquid spherical shells can form via a spinodal instability in the droplet center. A single liquid shell tends to grow until it undergoes a shape instability beyond a critical size. In an active emulsion, many stable and stationary liquid shells can coexist. We discuss conditions under which liquid shells are stable and dominant as compared to regimes where droplets undergo shape instabilities and divide.
Daxiao Sun, Xueping Zhao, Tina Wiegand, Giacomo Bartolucci, Cecilie Martin-Lemaitre, Stephan W. Grill, Anthony A. Hyman, Christoph Weber, Alf Honigmann
Formation of biomolecular condensates via phase separation enables compartmentation of many cellular processes. However, how cells can control condensation at specific locations to create complex cellular structures remains poorly understood. Here, we investigated the mechanism of tight junction formation, which involves condensation of scaffold proteins at cell-cell contacts and elongation of the condensates into a belt around the cellular perimeter. Using cell biology, reconstitution, and thermodynamic theory, we discovered that cells use surface phase transitions to control local condensation at the membrane far below bulk saturation. Surface condensation of junctional ZO-scaffold proteins is mediated by receptor binding and regulated by the receptor’s oligomerization state. Functionally, ZO surface condensation is directly coupled to actin polymerization and bundling, which drives elongation of receptor-ZO-actin condensates similar to tight junction belt formation in cells. We conclude that surface phase transitions provide a robust mechanism to control the position and shape of protein condensates.
Reyhaneh Khasseh, Sascha Wald, Roderich Moessner, Christoph A. Weber, Markus Heyl
Flocks of animals represent a fascinating archetype of collective behavior in the macroscopic classical world, where the constituents, such as birds, concertedly perform motions and actions as if being one single entity. Here, we address the outstanding question of whether flocks can also form in the microscopic world at the quantum level. For that purpose, we introduce the concept of active quantum matter by formulating a class of models of active quantum particles on a one-dimensional lattice. We provide both analytical and large-scale numerical evidence that these systems can give rise to quantum flocks. A key finding is that these flocks, unlike classical ones, exhibit distinct quantum properties by developing strong quantum coherence over long distances. We propose that quantum flocks could be experimentally observed in Rydberg atom arrays. Our work paves the way towards realizing the intriguing collective behaviors of biological active particles in quantum matter systems. We expect that this opens up a path towards a yet totally unexplored class of nonequilibrium quantum many-body systems with unique properties.
Jonathan Bauermann, Sudarshana Laha, Patrick M. McCall, Frank Jülicher, and Christoph A. Weber
The kinetics of chemical reactions are determined by the law of mass action, which has been successfully applied to homogeneous, dilute mixtures. At non-dilute conditions, interactions among the components can give rise to coexisting phases, which can significantly alter the kinetics of chemical reactions. Here, we derive a theory for chemical reactions in coexisting phases at phase equilibrium. We show that phase equilibrium couples the rates of chemical reactions of components with their diffusive exchanges between the phases. Strikingly, the chemical relaxation kinetics can be represented as a flow along the phase equilibrium line in the phase diagram. A key finding of our theory is that differences in reaction rates between coexisting phases stem solely from phase- dependent reaction rate coefficients. Our theory is key to interpret how concentration levels of reactive components in condensed phases control chemical reaction rates in synthetic and biological systems.
Thomas C.T. Michaels, L. Mahadevan and Christoph A. Weber
Liquid condensates are membraneless organelles that form via phase separation in living cells. These condensates provide unique heterogeneous environments that have much potential in regulating a range of biochemical processes from gene expression to filamentous protein aggregation—a process linked to Alzheimer's and Parkinson's diseases. Here we theoretically study the physical interplay between protein aggregation, its inhibition, and liquid-liquid phase separation. Our key finding is that the action of protein aggregation inhibitors can be strongly enhanced by liquid condensates. The physical mechanism of this enhancement relies on the partitioning and colocalization of inhibitors with their targets inside the liquid condensate. Our theory uncovers how the physicochemical properties of condensates can be used to modulate inhibitor potency, and we provide experimentally testable conditions under which drug potency is maximal. Our findings suggest design principles for protein aggregation inhibitors with respect to their phase-separation properties.
Joël Mabillard, Christoph A. Weber, Frank Jülicher
Chemically active systems such as living cells are maintained out of thermal equilibrium due to chemical events which generate heat and lead to active fluctuations. A key question is to understand on which time and length scales active fluctuations dominate thermal fluctuations. Here, we formulate a stochastic field theory with Poisson white noise to describe the heat fluctuations which are generated by stochastic chemical events and lead to active temperature fluctuations. We find that on large length and time scales, active fluctuations always dominate thermal fluctuations. However, at intermediate length and time scales, multiple crossovers exist which highlight the different characteristics of active and thermal fluctuations. Our work provides a framework to characterize fluctuations in active systems and reveals that local equilibrium holds at certain length and time scales.
Wolfram Pönisch, Thomas C.T. Michaels, and Christoph A. Weber
Biomolecular condensates in living cells can exhibit a complex rheology, including viscoelastic and glassy behavior. This rheological behavior of condensates was suggested to regulate polymerization of cytoskeletal filaments and aggregation of amyloid fibrils. Here, we theoretically investigate how the rheological properties of condensates can control the formation of linear aggregates. To this end, we propose a kinetic theory for linear aggregation in coexisting phases, which accounts for the aggregate size distribution and the exchange of aggregates between inside and outside of condensates. The rheology of condensates is accounted in our model via aggregate mobilities that depend on aggregate size. We show that condensate rheology determines whether aggregates of all sizes or dominantly small aggregates are exchanged between condensate inside and outside on the timescale of aggregation. As a result, the ratio of aggregate numbers inside to outside of condensates differs significantly. Strikingly, we also find that weak variations in the rheological properties of condensates can lead to a switch-like change of the number of aggregates. These results suggest a possible physical mechanism for how living cells could control linear aggregation in a switch-like fashion through variations in condensate rheology.
Jonathan Bauermann, Christoph A. Weber, Frank Jülicher
Chemically active droplets provide simple models for cell-like systems that can grow and divide. Such active droplet systems are driven away from thermodynamic equilibrium and turn over chemically, which corresponds to a simple metabolism. Two scenarios of nonequilibrium driving are considered. First, droplets are driven via the system boundaries by external reservoirs that supply nutrient and remove waste (boundary-driven). Second, droplets are driven by a chemical energy provided by a fuel in the bulk (bulk-driven). For both scenarios, the conservation of energy and matter as well as the balance of entropy are discussed. Conserved and nonconserved fields are used to analyse the nonequilibrium steady states of active droplets. Using an effective droplet model, droplet stability and instabilities leading to droplet division are explored. This work reveals that droplet division occurs quite generally in active droplet systems. The results suggest that life-like processes such as metabolism and division can emerge in simple nonequilibrium systems that combine the physics of phase separation and chemical reactions.
Tiemei Lu, Susanne Liese, Ludo Schoenmakers, Christoph A. Weber, Hiroaki Suzuki, Wilhelm T.S. Huck and Evan Spruijt
Recent studies have shown that the interactions between condensates and biological membranes are of functional importance. Here, we study how the interaction between complex coacervates and liposomes as model systems can lead to wetting, membrane deformation, and endocytosis. Depending on the interaction strength between coacervates and liposomes, the wetting behavior ranged from nonwetting to engulfment (endocytosis) and complete wetting. Endocytosis of coacervates was found to be a general phenomenon: coacervates made from a wide range of components could be taken up by liposomes. A simple theory taking into account surface energies and coacervate sizes can explain the observed morphologies. Our findings can help to better understand condensate–membrane interactions in cellular systems and provide new avenues for intracellular delivery using coacervates.
Xueping Zhao, Giacomo Bartolucci, Alf Honigmann, Frank Jülicher, Christoph A. Weber
In living cells, protein-rich condensates can wet the cell membrane and surfaces of membrane-bound organelles. Interestingly, many phase-separating proteins also bind to membranes leading to a molecular layer of bound molecules. Here we investigate how binding to membranes affects wetting, prewetting and surface phase transitions. We derive a thermodynamic theory for a three-dimensional bulk in the presence of a two-dimensional, flat membrane. At phase coexistence, we find that membrane binding facilitates complete wetting and thus lowers the wetting angle. Moreover, below the saturation concentration, binding facilitates the formation of a thick layer at the membrane and thereby shifts the prewetting phase transition far below the saturation concentration. The distinction between bound and unbound molecules near the surface leads to a large variety of surface states and complex surface phase diagrams with a rich topology of phase transitions. Our work suggests that surface phase transitions combined with molecular binding represent a versatile mechanism to control the formation of protein-rich domains at intra-cellular surfaces.
Stefano Bo, Lars Hubatsch, Jonathan Bauermann, Christoph A. Weber, and Frank Jülicher
We discuss the stochastic trajectories of single molecules in a phase-separated liquid, when a dense and a dilute phase coexist. Starting from a continuum theory of macroscopic phase separation we derive a stochastic Langevin equation for molecular trajectories that takes into account thermal fluctuations. We find that molecular trajectories can be described as diffusion with drift in an effective potential, which has a steep gradient at phase boundaries. We discuss how the physics of phase coexistence affects the statistics of molecular trajectories and in particular the statistics of displacements of molecules crossing a phase boundary. At thermodynamic equilibrium detailed balance imposes that the distributions of displacements crossing the phase boundary from the dense or from the dilute phase are the same. Our theory can be used to infer key phase separation parameters from the statistics of single-molecule trajectories. For simple Brownian motion, there is no drift in the presence of a concentration gradient. We show that interactions in the fluid give rise to an average drift velocity in concentration gradients. Interestingly, under non-equilibrium conditions, single molecules tend to drift uphill the concentration gradient. Thus, our work bridges between single-molecule dynamics and collective dynamics at macroscopic scales and provides a framework to study single-molecule dynamics in phase-separating systems.
Giacomo Bartolucci, Omar Adame-Arana, Xueping Zhao, and Christoph A. Weber
Phase separation and transitions among different molecular states are ubiquitous in living cells. Such transitions can be governed by local equilibrium thermodynamics or by active processes controlled by biological fuel. It remains largely unexplored how the behavior of phase-separating systems with molecular transitions differs between thermodynamic equilibrium and cases in which the detailed balance of the molecular transition rates is broken because of the presence of fuel. Here, we present a model of a phase-separating ternary mixture in which two components can convert into each other. At thermodynamic equilibrium, we find that molecular transitions can give rise to a lower dissolution temperature and thus reentrant phase behavior. Moreover, we find a discontinuous thermodynamic phase transition in the composition of the droplet phase if both converting molecules attract themselves with similar interaction strength. Breaking the detailed balance of the molecular transition leads to quasi-discontinuous changes in droplet composition by varying the fuel amount for a larger range of intermolecular interactions. Our findings showcase that phase separation with molecular transitions provides a versatile mechanism to control properties of intracellular and synthetic condensates via discontinuous switches in droplet composition.
Lars Hubatsch, Louise M. Jawerth, Celina Love, Jonathan Bauermann, T-Y Dora Tang, Stefano Bo, Anthony A. Hyman, and Christoph A. Weber
Key processes of biological condensates are diffusion and material exchange with their environment. Experimentally, diffusive dynamics are typically probed via fluorescent labels. However, to date, a physics-based, quantitative framework for the dynamics of labeled condensate components is lacking. Here, we derive the corresponding dynamic equations, building on the physics of phase separation, and quantitatively validate the related framework via experiments. We show that by using our framework, we can precisely determine diffusion coefficients inside liquid condensates via a spatio-temporal analysis of fluorescence recovery after photobleaching (FRAP) experiments. We showcase the accuracy and precision of our approach by considering space- and time-resolved data of protein condensates and two different polyelectrolyte-coacervate systems. Interestingly, our theory can also be used to determine a relationship between the diffusion coefficient in the dilute phase and the partition coefficient, without relying on fluorescence measurements in the dilute phase. This enables us to investigate the effect of salt addition on partitioning and bypasses recently described quenching artifacts in the dense phase. Our approach opens new avenues for theoretically describing molecule dynamics in condensates, measuring concentrations based on the dynamics of fluorescence intensities, and quantifying rates of biochemical reactions in liquid condensates.
Local thermodynamics govern formation and dissolution of Caenorhabditis elegans P granule condensates
Anatol W. Fritsch, Andrés F. Diaz-Delgadillo, Omar Adame-Arana, Carsten Hoege, Matthäus Mittasch, Moritz Kreysing, Mark Leaver, Anthony A. Hyman, Frank Jülicher, Christoph A. Weber
Membraneless compartments, also known as condensates, provide chemically distinct environments and thus spatially organize the cell. A well-studied example of condensates is P granules in the roundworm Caenorhabditis elegans that play an important role in the development of the germline. P granules are RNA-rich protein condensates that share the key properties of liquid droplets such as a spherical shape, the ability to fuse, and fast diffusion of their molecular components. An outstanding question is to what extent phase separation at thermodynamic equilibrium is appropriate to describe the formation of condensates in an active cellular environment. To address this question, we investigate the response of P granule condensates in living cells to temperature changes. We observe that P granules dissolve upon increasing the temperature and recondense upon lowering the temperature in a reversible manner. Strikingly, this temperature response can be captured by in vivo phase diagrams that are well described by a Flory–Huggins model at thermodynamic equilibrium. This finding is surprising due to active processes in a living cell. To address the impact of such active processes on intracellular phase separation, we discuss temperature heterogeneities. We show that, for typical estimates of the density of active processes, temperature represents a well-defined variable and that mesoscopic volume elements are at local thermodynamic equilibrium. Our findings provide strong evidence that P granule assembly and disassembly are governed by phase separation based on local thermal equilibria where the nonequilibrium nature of the cytoplasm is manifested on larger scales.
Patrick Schwarz, Sudarshana Laha, Jacqueline Janssen, Tabea Huss, Job Boekhoven, Christoph A. Weber
Non-equilibrium, fuel-driven reaction cycles serve as model systems of the intricate reaction networks of life. Rich and dynamic behavior is observed when reaction cycles regulate assembly processes, such as phase separation. However, it remains unclear how the interplay between multiple reaction cycles affects the success of emergent assemblies. To tackle this question, we created a library of molecules that compete for a common fuel that transiently activates products. Often, the competition for fuel implies that a competitor decreases the lifetime of these products. However, in cases where the transient competitor product can phase-separate, such a competitor can increase the survival time of one product. Moreover, in the presence of oscillatory fueling, the same mechanism reduces variations in the product concentration while the concentration variations of the competitor product are enhanced. Like a parasite, the product benefits from the protection of the host against deactivation and increases its robustness against fuel variations at the expense of the robustness of the host. Such a parasitic behavior in multiple fuel-driven reaction cycles represents a lifelike trait, paving the way for the bottom-up design of synthetic life.
Drops in cells: Liquid droplets act as microreactors. Can they also serve as a control mechanism for cellular biochemistry?
Christoph A. Weber and Christoph Zechner
A major challenge in cell biology remains unraveling how cells control their biochemical reaction cycles. For instance, how do they regulate gene expression in response to stress? How does their metabolism change when resources are scarce? Control theory has proven useful in understanding how networks of chemical reactions can robustly tackle those and other tasks.1 The essential ingredients in such approaches are chemical feedback loops that create control mechanisms similar to the circuits that regulate, for example, the temperature of a heating system, the humidity of an archive, or the pH of a fermentation tank.
Christoph A. Weber
The physics of phase separation and the formation of protein-rich droplets play an important role for biochemical processes in living cells. The shape, size and composition of such droplets can change with time and thereby affect biochemical reactions. Such reactions also affect the dynamics of droplets. Obtaining insights into this interplay is key to better understand the mechanisms underlying the spatio-temporal organization of biological cells.
Christoph A. Weber, Lars Hubatsch, Frank Jülicher
Zellen führen biochemische Prozesse aus, um zentrale Abläufe wie die Zellteilung zu realisieren. Bei der dafür erforderlichen raumzeitlichen Organisation der zellulären Prozesse kommt den Organellen eine wichtige Rolle zu. Organellen wie die Mitochondrien oder der Zellkern sind durch Membranen von ihrer Umgebung getrennt. Diese ermöglichen es ihnen, in ihrem Inneren geeignete biochemische Bedingungen für biologische Prozesse zu erzeugen. Doch es gibt auch membranlose Organellen. Wie bewahren diese ihre chemische Identität? Hierbei kommt die Koexistenz proteinreicher, flüssiger Phasen ins Spiel. Ausgehend von der Physikder Phasenseparation ist ein tieferes Verständnis der Dynamik und raumzeitlichen Organisation biochemischer Prozesse möglich.
Omar Adame-Arana, Christoph A. Weber, Vasily Zaburdaev, Jacques Prost, Frank Jülicher
We present a minimal model to study the effects of pH on liquid phase separation of macromolecules. Our model describes a mixture composed of water and macromolecules that exist in three different charge states and have a tendency to phase separate. This phase separation is affected by pH via a set of chemical reactions describing protonation and deprotonation of macromolecules, as well as self-ionization of water. We consider the simple case in which interactions are captured by Flory-Huggins interaction parameters corresponding to Debye screening lengths shorter than a nanometer, which is relevant to proteins inside biological cells under physiological conditions. We identify the conjugate thermodynamic variables at chemical equilibrium and discuss the effective free energy at fixed pH. First, we study phase diagrams as a function of macromolecule concentration and temperature at the isoelectric point of the macromolecules. We find a rich variety of phase diagram topologies, including multiple critical points, triple points, and first-order transition points. Second, we change the pH relative to the isoelectric point of the macromolecules and study how phase diagrams depend on pH. We find that these phase diagrams as a function of pH strongly depend on whether oppositely charged macromolecules or neutral macromolecules have a stronger tendency to phase separate. One key finding is that we predict the existence of a reentrant behavior as a function of pH. In addition, our model predicts that the region of phase separation is typically broader at the isoelectric point. This model could account for both in vitro phase separation of proteins as a function of pH and protein phase separation in yeast cells for pH values close to the isoelectric point of many cytosolic proteins.
Marta Tena-Solsona, Jacqueline Janssen, Caren Wanzke, Fabian Schnitter, Hansol Park, Benedikt Rieß, Julianne M. Gibbs, Christoph A. Weber, Job Boekhoven
Chemically fueled emulsions are solutions with droplets made of phase-separated molecules that are activated and deactivated by a chemical reaction cycle. These emulsions play a crucial role in biology as a class of membrane-less organelles. Moreover, theoretical studies show that droplets in these emulsions can evolve to the same size or spontaneously self-divide when fuel is abundant. All of these exciting properties, i. e., emergence, decay, collective behavior, and self-division, are pivotal to the functioning of life. However, these theoretical predictions lack experimental systems to test them quantitively. Here, we describe the synthesis of synthetic emulsions formed by a fuel-driven chemical cycle, and we find a surprising new behavior, i. e., the dynamics of droplet growth is regulated by the kinetics of the fuel-driven reaction cycle. Consequently, the average volume of these droplets grows orders of magnitude faster compared to Ostwald ripening. Combining experiments and theory, we elucidate the underlying mechanism.
Nicholas J. Derr, David C. Fronk, Christoph A. Weber, Amala Mahadevan, Chris H. Rycroft, L. Mahadevan
Channel formation and branching is widely seen in physical systems where movement of fluid through a porous structure causes the spatiotemporal evolution of the medium. We provide a simple theoretical framework that embodies this feedback mechanism in a multiphase model for flow through a frangible porous medium with a dynamic permeability. Numerical simulations of the model show the emergence of branched networks whose topology is determined by the geometry of external flow forcing. This allows us to delineate the conditions under which splitting and/or coalescing branched network formation is favored, with potential implications for both understanding and controlling branching in soft frangible media.
Thomas CT. Michaels, Christoph A. Weber, L Mahadevan
Protein aggregation has been implicated in many medical disorders, including Alzheimer’s and Parkinson’s diseases. Potential therapeutic strategies for these diseases propose the use of drugs to inhibit specific molecular events during the aggregation process. However, viable treatment protocols require balancing the efficacy of the drug with its toxicity, while accounting for the underlying events of aggregation and inhibition at the molecular level. To address this key problem, we combine here protein aggregation kinetics and control theory to determine optimal protocols that prevent protein aggregation via specific reaction pathways. We find that the optimal inhibition of primary and fibril-dependent secondary nucleation require fundamentally different drug administration protocols. We test the efficacy of our approach on experimental data for the aggregation of the amyloid-β(1-42) peptide of Alzheimer’s disease in the model organism Caenorhabditis elegans. Our results pose and answer the question of the link between the molecular basis of protein aggregation and optimal strategies for inhibiting it, opening up avenues for the design of rational therapies to control pathological protein aggregation.
Christoph A. Weber, David Zwicker, Frank Jülicher and Chiu Fan Lee
Phase separating systems that are maintained away from thermodynamic equilibrium via molecular processes represent a class of active systems, which we call active emulsions. These systems are driven by external energy input, for example provided by an external fuel reservoir. The external energy input gives rise to novel phenomena that are not present in passive systems. For instance, concentration gradients can spatially organise emulsions and cause novel droplet size distributions. Another example are active droplets that are subject to chemical reactions such that their nucleation and size can be controlled, and they can divide spontaneously. In this review, we discuss the physics of phase separation and emulsions and show how the concepts that govern such phenomena can be extended to capture the physics of active emulsions. This physics is relevant to the spatial organisation of the biochemistry in living cells, for the development of novel applications in chemical engineering and models for the origin of life.
Christoph A. Weber, Thomas C. T. Michaels, L. Mahadevan
Liquid cellular compartments form in the cyto- or nucleoplasm and can regulate aberrant protein aggregation. Yet, the mechanisms by which these compartments affect protein aggregation remain unknown. Here, we combine kinetic theory of protein aggregation and liquid-liquid phase separation to study the spatial control of irreversible protein aggregation in the presence of liquid compartments. We find that even for weak interactions aggregates strongly partition into the liquid compartment. Aggregate partitioning is caused by a positive feedback mechanism of aggregate nucleation and growth driven by a flux maintaining the phase equilibrium between the compartment and its surrounding. Our model establishes a link between specific aggregating systems and the physical conditions maximizing aggregate partitioning into the compartment. The underlying mechanism of aggregate partitioning could be used to confine cytotoxic protein aggregates inside droplet-like compartments but may also represent a common mechanism to spatially control irreversible chemical reactions in general.
Wolfram Pönisch, Christoph A. Weber, Vasily Zaburdaev
Many bacteria rely on active cell appendages, such as type IV pili, to move over substrates and interact with neighboring cells. Here, we study the motion of individual cells and bacterial colonies, mediated by the collective interactions of multiple pili. It was shown experimentally that the substrate motility of Neisseria gonorrhoeae cells can be described as a persistent random walk with a persistence length that exceeds the mean pili length. Moreover, the persistence length increases for a higher number of pili per cell. With the help of a simple, tractable stochastic model, we test whether a tug of war without directional memory can explain the persistent motion of single Neisseria gonorrhoeae cells. While persistent motion of single cells indeed emerges naturally in the model, a tug of war alone is not capable of explaining the motility of microcolonies, which becomes weaker with increasing colony size. We suggest sliding friction between the microcolonies and the substrate as the missing ingredient. While such friction almost does not affect the general mechanism of single cell motility, it has a strong effect on colony motility. We validate the theoretical predictions by using a three-dimensional computational model that includes explicit details of the pili dynamics, force generation, and geometry of cells.