Out-of-equilibrium self-assembly of proteins such as actin and tubulin is a key regulatory process controlling cell shape, motion and division. The design of functional nanosystems based on dissipative self-assembly has proven to be remarkably difficult due to a complete lack of control over the spatial and temporal characteristics of the assembly process. Here, we show the dissipative self-assembly of FtsZ protein (a bacterial homologue of tubulin) within coacervate droplets. More specifically, we show how such barrier-free compartments govern the local availability of the energy-rich building block guanosine triphosphate, yielding highly dynamic fibrils. The increased flux of FtsZ monomers at the tips of the fibrils results in localized FtsZ assembly, elongation of the coacervate compartments, followed by division of the fibrils into two. We rationalize the directional growth and division of the fibrils using dissipative reaction-diffusion kinetics and capillary action of the filaments as main inputs. The principle presented here, in which open compartments are used to modulate the rates of dissipative self-assembly by restricting the absorption of energy from the environment, may provide a general route to dissipatively adapting nanosystems exhibiting life-like behaviour.
The cytosol of Escherichia coli is an extremely crowded environment, containing high concentrations of biopolymers which occupy 20-30% of the available volume. Such conditions are expected to yield depletion forces, which strongly promote macromolecular complexation. However, crowded macromolecule solutions, like the cytosol, are very prone to nonspecific associative interactions that can potentially counteract depletion. It remains unclear how the cytosol balances these opposing interactions. We used a FRET-based probe to systematically study depletion in vitro in different crowded environments, including a cytosolic mimic, E. coli lysate. We also studied bundle formation of FtsZ protofilaments under identical crowded conditions as a probe for depletion interactions at much larger overlap volumes of the probe molecule. The FRET probe showed a more compact conformation in synthetic crowding agents, suggesting strong depletion interactions. However, depletion was completely negated in cell lysate and other protein crowding agents, where the FRET probe even occupied slightly more volume. In contrast, bundle formation of FtsZ protofilaments proceeded as readily in E. coli lysate and other protein solutions as in synthetic crowding agents. Our experimental results and model suggest that, in crowded biopolymer solutions, associative interactions counterbalance depletion forces for small macromolecules. Furthermore, the net effects of macromolecular crowding will be dependent on both the size of the macromolecule and its associative interactions with the crowded background.
We report a microfluidic approach to generate aqueous droplets in oil of different dimensionality, stabilized by a lipid monolayer, to systematically probe the polymerization of bacterial cell-division protein FtsZ into fibrous networks as a function of the concentrations of crowding agent, FtsZ, and GTP.FtsZ bundles confined in droplets were dynamic, and their distribution depended on the intrinsic properties of the system and restrictions imposed by the spatial boundaries. § Corresponding authors: Wilhelm T. S. Huck,
The versatility of polyelectrolyte multilayer (PEM) coatings is very promising for their use as separation layers in nanofiltration applications. These membranes can for example be suited for the removal of micropollutants, such as medicines and pesticides, from water. The selectivity of PEM coatings can be further improved by so-called asymmetric coating. In this approach, the pores of the support membrane are filled with an open PEM layer to maintain a good water permeability, and subsequently a thin, dense layer is coated on top to determine the separation properties. Coating a dense top layer can be achieved in different ways. In this work, we systematically investigate the effectiveness of these different types of top layers. We show that coating a top layer at lower ionic strength than the bottom layer leads to a higher permeability and MgSO4 retention, compared with the reference, bottom-type layer coated with the same total number of layers. Also, other salt retentions can be improved with this approach. However, micropollutant retentions are hardly affected. Coating a top layer with a polyelectrolyte pair that forms denser layers at equal ionic strength, in contrast, leads to a significant change in separation properties with much higher MgSO4 and micropollutant retentions and improved water permeability compared with the reference layer. The concept of membrane optimization via asymmetric coating is thus most effective when using different polyelectrolyte pairs on top of each other. Moreover, we show that this approach allows us to selectively cross-link the top layer for further enhancement of the micropollutant retention, while water permeability is not much reduced. Asymmetric PEM coatings are therefore a promising method to optimize PEM membranes for micropollutant removal and other separation processes.
Glucosinolates are secondary plant compounds typically found in members of the Brassicaceae and a few other plant families. Usually each plant species contains a specific subset of the ∼ 130 different glucosinolates identified to date. However, intraspecific variation in glucosinolate profiles is commonly found. Sinalbin (4-hydroxybenzyl glucosinolate) so far has been identified as the main glucosinolate of the heavy metal accumulating plant species Noccaea caerulescens (Brassicaceae). However, a screening of 13 N. caerulescens populations revealed that in 10 populations a structurally related glucosinolate was found as the major component. Based on nuclear magnetic resonance (NMR) and mass spectrometry analyses of the intact glucosinolate as well as of the products formed after enzymatic conversion by sulfatase or myrosinase, this compound was identified as 4-α-rhamnosyloxy benzyl glucosinolate (glucomoringin). So far, glucomoringin had only been reported as the main glucosinolate of Moringa spp. (Moringaceae) which are tropical tree species. There was no apparent relation between the level of soil pollution at the location of origin, and the presence of glucomoringin. The isothiocyanate that is formed after conversion of glucomoringin is a potent antimicrobial and antitumor agent. It has yet to be established whether glucomoringin or its breakdown product have an added benefit to the plant in its natural habitat.
The increase of micropollutant concentration in both surface and groundwater is an emerging concern for the environment and human health. Most of such small organic molecules (medicines, hormones, and plasticizers) enter the environment via our wastewater, because they are not sufficiently removed by the current techniques applied in wastewater treatment plants. A possible solution to remove micropollutants is the usage of polyelectrolyte multilayer (PEM) based membranes. PEM membranes have received a growing interest in the past decade due to their high chemical and physical stability and their high permeability and selectivity. A popular polyelectrolyte pair to make dense PEM membranes with high salt retentions is the combination of poly(allylamine hydrochloride) (PAH) and poly(sodium 4-styrenesulfonate) (PSS). Unfortunately, smaller micropollutants (such as bisphenol A, sulfamethoxazole, naproxen, and bezafibrate) still show significant permeation through this membrane. In this study, for the first time, a single final layer of Nafion is applied on the PEM to increase the density of the PEM membrane. It is shown that when terminating with Nafion, the swelling of the multilayer decreases by 50%. These pronounced changes in layer structure are reflected by changes in membrane performance, such as a lower molecular weight cutoff (MWCO) and an increasing hydraulic membrane resistance. Furthermore, we show that the Nafion content of the multilayer can be increased by constructing a Nafion/PAH multilayer on top of the existing PSS/PAH multilayer, thereby lowering the MWCO. Although hydraulic resistance increases, these PSS/PAH/Nafion-based multilayers show excellent performance in rejecting difficult-to-remove micropollutants that have low molecular weight (200–650 Da) and different charges. Overall, a cocktail of eight small micropollutants can be removed up to 97% by these membranes, allowing strongly enhanced water purification.
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