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Eumelanin exhibits a defined supramolecular buildup that is deprived of at least three distinct particle species. To enable the full potential of its promising material properties, access to all particle types is crucial. In this work, the first protocol for the synthesis of the intermediate type-A particles in pure and stable dispersion form is described. It is found that aggregation of type-A particles into the larger type-B variant can be inhibited by a strict pH control during the synthesis. The exact influence of pH on the supramolecular buildup is investigated via a combination of time-resolved light scattering, electron microscopy, and UV-vis spectroscopy. It is observed that a rapid buildup of type-B particles occurs without pH control and is generally dominant at lower pH values. At pH values above 6.2 however, type-A particles are gained, and no further aggregation occurs. Even more, lowering the pH of such a stable type-A dispersion at a later stage lifts the inhibition and again leads to the formation of larger particle species. The results confirm that it is easily possible to halt the aggregation of eumelanin substructures and to access them in the form of a stable dispersion. Moreover, a profound additional understanding of the supramolecular buildup is gained by the in-depth investigation of the pH influence.

The natural blood protein fibrinogen is a highly potent precursor for the production of various biomaterials due to its supreme biocompatibility and cell interaction. To gain actual materials from fibrinogen, the protein needs to undergo fibrillogenesis, which is mostly triggered via enzymatic processing to fibrin, electrospinning, or drying processes. All of those techniques, however, strongly limit the available structures or the applicability of the material. To overcome the current issues of fibrin(ogen) as material, we herein present a highly feasible, quick, and inexpensive technique for self-assembly of fibrinogen in solution into defined, nanofibrous three-dimensional (3D) patterns. Upon interaction with specific anions in controlled environments, stable and flexible hydrogel-like structures are formed without any further processing. Moreover, the material can be converted into highly porous and elastic aerogels by lyophilization. Both of these material classes have never been described before from native fibrinogen. The observed phenomenon also represents the first enzyme-free process of fibrillogenesis from fibrinogen with significant yield in solution. The produced hydrogels and aerogels were investigated via electron microscopy, IR spectroscopy, and fluorescence spectroscopy, which also confirms the native state of the protein. Additionally, their mechanical properties were compared with actual fibrin and unstructured fibrinogen. The structural features show a striking analogy to actual fibrin, both as hydro- and aerogel. This renders the new material a highly promising alternative for fibrin in biomaterial applications. A much faster initiation of fiber formation, exclusion of possible thrombin residuals, and low-cost reagents are great advantages.

A disulfide polymer that is triggerable by ultrasound, UV, and redox in aqueous solution and can self-assemble with ionic probe molecules into again multitriggerable assemblies is presented. It is ...

Pseudo isocyanine chloride (PIC) has a strong tendency to self-assemble with a concentration dependent temperature threshold separating a regime with small H-oligomers in equilibrium with monomeric PIC from a regime where large J-aggregates form. In complementing the known set of absorption spectra by the spectrum of a trimer, which represents all H-aggregates with N ≥ 3, a full description of the sample composition of PIC in the regime of oligomers became possible by means of UV-vis spectroscopy and gave access to the equilibrium thermodynamics of oligomerisation. Successful interpretation of the concentration dependent temperature threshold as a ceiling temperature of J-aggregation made also accessible equilibrium thermodynamics of the formation of J-aggregates together with a full analysis of composition also in the regime of J-aggregates. Evolution of an invariant spectrum for the J-aggregates demonstrates full consistency of the composition analysis. Complementary light scattering experiments led to a comprehensive characterisation of all aggregate species above and below the aggregation threshold. Once initiated, J-aggregates always grow to a final length of 650 nm. Time-resolved light scattering confirmed that the self-assembly of J-aggregates follows a monomer addition process in analogy to a chain growth in polymer chemistry. Initiation and growth of individual aggregates turned out to be always much faster than the progress of aggregation.

Pseudo‐isocyanine chloride (PIC) is a cationic dyestuff that exhibits self‐assembly in aqueous solution, promoted either by increasing the PIC concentration or by decreasing the temperature. PIC‐aggregates exhibit a characteristic and sharp absorption band as well as a fluorescence band at a wavelength of 573 nm making PIC an interesting candidate to analyze the self‐assembly process in various environments. The present work developed PIC‐based, synthetic model systems, suitable to investigate how macromolecular crowding influences self‐assembly processes. Four synthetic additives were used as potential crowders: Triethylene glycol (TEG), polyethylene glycol (PEG), Ficoll 400 as a highly branched polysaccharide, and sucrose corresponding to the monomeric unit of Ficoll. Combined UV/Vis spectroscopy and time‐resolved light scattering revealed a strong impact of crowding based on excluded volume effects only for Ficoll 400. Sucrose had hardly any influence on the self‐assembly of PIC and PEG and TEG impeded the PIC self‐assembly. Development of such a PIC based model system led over to in‐cell experiments. HeLa cells were infiltrated with PIC solutions well below the aggregation threshold in the infiltrating solution. In the cellular environment, PIC was exposed to a significant crowding and immediately started to aggregate. As was demonstrated by fluorescence imaging, the extent of aggregation can be modulated by exposing the cells to salt‐induced osmotic stress. The results suggest future use of such a system as a sensor for the analysis of in vitro and in vivo crowding effects on self‐assembly processes., Stand out from the crowd! The cationic dyestuff PIC reversibly self‐assembles in aqueous solution. A comparative investigation of such PIC self‐assembly in the presence of synthetic polymers with PIC self‐assembly in living Hela cells suggests PIC as a promising candidate for a sensor to examine the impact of varying cellular conditions like macromolecular crowding on self‐assembly processes of proteins in cells (see figure).

Block copolymers were prepared with two anionic polyelectrolyte blocks: sodium polyacrylate (PA) and sodium polystyrene sulfonate (PSS), in order to investigate their phase behavior in aqueous solution in the presence of Ca2+ cations. Depending on the concentration of polymer and Ca2+ and on the ratio of the block lengths in the copolymer, spherical micelles were observed. Micelle formation arises from the specific interaction of Ca2+ with the PA block only. An extensive small-angle scattering study was performed in order to unravel the structure and dimensions of the block copolymer micelles. Deuteration of the PA block enabled us to perform contrast variation experiments using small-angle neutron scattering at variable ratios of light and heavy water which were combined with information from small-angle X-ray scattering and dynamic light scattering.

ChemistryOpen 11(4), e202200024 (2022). doi:10.1002/open.202200024, Protein aggregation is a hallmark of several severe neurodegenerative disorders such as Huntington's, Parkinson's, or Alzheimer's disease. Metal ions play a profound role in protein aggregation and altered metal-ion homeostasis is associated with disease progression. Here we utilize μ-X-ray fluorescence imaging in combination with rapid freezing to resolve the elemental distribution of phosphorus, sulfur, potassium, and zinc in huntingtin exon-1-mYFP expressing HeLa cells. Using quantitative XRF analysis, we find a threefold increase in zinc and a 10-fold enrichment of potassium that can be attributed to cellular stress response. While the averaged intracellular ion areal masses are significantly different in aggregate-containing cells, a local intracellular analysis shows no different ion content at the location of intracellular inclusion bodies. The results are compared to corresponding experiments on HeLa cells forming pseudoisocyanine chloride aggregates. As those show similar results, changes in ion concentrations are not exclusively linked to huntingtin exon-1 amyloid formation., Published by Wiley-VCH-Verl., Weinheim

Small angle light scattering (SALS) and differential scanning calorimetry DSC have been applied to investigate the melting of spherulites isothermally crystallized polylactide. At an isothermal crystallization temperature high enough, pure α‐phase crystals are formed. Exposed to a temperature gradient, the crystals first melt and then recrystallize before they finally melt. With decreasing crystallization temperature, an increasing fraction of polylactide is crystallizing in the less stable α ′‐phase. α ′‐crystals also melt upon increasing the temperature but recrystallize to the more stable α‐phase. A constant spherulite size is revealed by SALS for both processes, the α/α and α ′/α melt‐recrystallization, until completion of the final melting, thereby supporting integrity of the spherulites throughout the entire processes. Joint DSC and SALS experiments demonstrate that the depolarized scattering invariant correlates with the heat flow recorded by DSC and thus offer an alternative measure for the degree of crystallinity. The following mechanism is identified for both processes: initial melting and recrystallization overlay each other. Crystallinity is not fully recovered upon recrystallization because only part of the original lamellae survives the melt‐recrystallization, though with an increased thickness. While lamellae are melting and reforming or simply transforming their phase, the spherulites survive the process until final melting. © 2019 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2019, 57, 1483–1495

Zeolitic imidazolate frameworks (ZIFs) represent an important subclass of porous metal organic frameworks (MOFs). The present work continues a previous study on ZIF-71 nanocrystal formation at room temperature (Cryst. Growth Des. 2016, 16, 2002–2010), which proposed a four-step formation process. It introduces a kinetic nucleation and growth model, which satisfactorily reproduces in situ time-resolved static light scattering data measured at room temperature in the previous work and at variable temperatures in the present work. Successful fitting can only be achieved with a precursor reaction being put before particle nucleation. The kinetic analysis provides rate constants for the precursor reaction, for particle nucleation, and for particle growth via monomer addition, and the equilibrium concentration of nonconsumed matter at the end of reaction. The rate constants of the precursor reaction and the nucleation process increase with temperature, which is counterintuitive if compared to classical nucleati...

Complexes between the anionic polyelectrolyte sodium polyacrylate (PA) and an oppositely charged divalent azobenzene dye are prepared in aqueous solution. Depending on the ratio between dye and polyelectrolyte stable aggregates with a well-defined spherical shape are observed. Upon exposure of these complexes to UV light, the trans → cis transition of the azobenzene is excited resulting in a better solubility of the dye and a dissolution of the complexes. The PA chains reassemble into well-defined aggregates when the dye is allowed to relax back into the trans isomer. Varying the temperature during this reformation step has a direct influence on the final size of the aggregates rendering temperature in an efficient way to easily change the size of the self-assemblies. Application of time-resolved small-angle neutron scattering (SANS) to study the structure formation reveals that the cis → trans isomerization is the rate-limiting step followed by a nucleation and growth process.

The human guanylate-binding protein 1 (hGBP1) belongs to the dynamin superfamily proteins and represents a key player in the innate immune response. Farnesylation at the C-terminus is required for hGBP1's activity against microbial pathogens, as well as for its antiproliferative and antitumor activity. The farnesylated hGBP1 (hGBP1fn) retains many characteristics of the extensively studied nonfarnesylated protein and gains additional abilities like binding to lipid membranes and formation of hGBP1fn polymers. These polymers are believed to serve as a protein depot, making the enzyme immediately available to fight the invasion of intracellular pathogens. Here we study the molecular mechanism of hGBP1 polymer formation as it is a crucial state of this enzyme, allowing for a rapid response demanded by the biological function. We employ Forster resonance energy transfer in order to trace intra and intermolecular distance changes of protein domains. Light scattering techniques yield deep insights into the changes in size and shape. The GTP hydrolysis driven cycling between a closed, farnesyl moiety hidden state and an opened, farnesyl moiety exposed state represents the first phase, preparing the molecule for polymerization. Within the second phase of polymer growth, opened hGBP1 molecules can be incorporated in the growing polymer where the opened structure is stabilized, similar to a surfactant molecule in a micelle, pointing the farnesyl moieties into the hydrophobic center and positioning the head groups at the periphery of the polymer. We contribute the molecular mechanism of polymer formation, paving the ground for a detailed understanding of hGBP1 function.

The interior of a cell is a highly packed environment that can be occupied up to 40% by different macromolecules. Such crowded media influence different biochemical processes like protein folding, enzymatic activity, and gene regulation. In this work, we use simulations to study protein stability under the presence of crowding agents that interact with the protein by excluded volume interactions. In general, the presence of crowding agents in the solution enhances the stability of the protein's native state. However, we find that the effects of excluded volume depend not only on crowding occupancy but also the crowders' geometry and size. Specifically, we find that polymeric crowders have stronger influence than spherical crowders and that this effect increases with polymer length, while it decreases with increasing size of spherical crowders. These opposing size effects are explained by the interplay of decreasing excluded volume and demixing, which together determine the change in the entropy of the crowders upon folding of the protein.

Fibrinogen not only forms fibrin networks if assisted by thrombin but also exhibits self-assembly in dilute aqueous solutions in the absence of thrombin. It could be shown that self-assembly can be triggered in a controlled way by diluting the ionic strength set to a value of 0.14 M NaCl in the starting solutions. The present work unravels the mechanism of this self-assembly process by means of a combination of time-resolved multiangle static and dynamic light scattering and atomic force microscopy. Analysis was carried out as a function of the ionic strength adjusted by the drop in ionic strength and at variable salt compositions at a given final ionic strength. Composition was varied by changing the ratio of NaCl and phosphate buffer. The self-assembly induced by the drop of the ionic strength depends on the final value. The lower the final ionic strength gets, the faster is the self-assembly process. The variation of the salt composition at a given ionic strength has only a marginal effect, which depends on the ionic strength. The self-assembly obeys a step-growth process, where any intermediate cluster can coalesce with any other cluster. Interpretation of the data with a kinetic model based on the approach of von Smoluchowski follows a diffusion-limited cluster aggregation at ionic strength values lower than 30 mM. At an ionic strength of 30 mM, the model has to take into account a size dependence of the rate constant, and at 60 mM a transition is observed to a reaction-limited cluster aggregation.

Unlike Na+ cations, which interact with fully neutralized poly(acrylic acid) as purely electrostatic entities, cations like Ag+, Mg2+, and Ca2+ exhibit specific interactions with the COO– residues of anionic polyacrylates. The present work analyzes the interaction of all four cations with a layer of polyacrylate chains grafted onto spherical polystyrene core as an outer shell. First and foremost the analysis answers the question on how these specific interactions influence the nature of the shell and solution behavior of the spherical polyelectrolyte brushes. It could be unambiguously demonstrated that Ag+, Mg2+, and Ca2+ cations induce a drastic shrinking of the polyacrylate shells at cation concentrations lower by 2–3 orders of magnitude compared to the transition of a fully stretched osmotic brush to a shrunken salted brush accomplished with Na+ cations. Ag+ induces a particularly abrupt contraction, which comes close to the annealing of such brushes achieved with a neutralization by protons. Finally, ...

Vimentin intermediate filaments (IFs) are part of a family of proteins that constitute one of the three filament systems in the cytoskeleton, a major contributor to cell mechanics. One property that distinguishes IFs from the other cytoskeletal filament types, actin filaments and microtubules, is their highly hierarchical assembly pathway, where a lateral association step is followed by elongation. Here we present an innovative technique to follow the elongation reaction in solution and in situ by time-resolved static and dynamic light scattering, thereby precisely capturing the relevant time and length scales of seconds to minutes and 60–600 nm, respectively. We apply a quantitative model to our data and succeed in consistently describing the entire set of data, including particle mass, radius of gyration, and hydrodynamic radius during longitudinal association.

The current rather poor understanding of the mechanisms of crystallization of zeolitic imidazolate frameworks (ZIFs), an important subclass of porous metal organic framework (MOF) materials, greatly hampers rational synthesis of predicted ZIF phases/polymorphs and control of crystal size/shape. In this contribution, we present the results of in situ studies on a recently developed fast ZIF-71 nanocrystal synthesis from solution. By taking advantage of the combined use of time-resolved static and dynamic light scattering and stopped-flow synchrotron small-angle and wide-angle X-ray scattering, we were able to reveal the whole nanocrystal formation process: a population of small amorphous particles (termed clusters) first forms via coagulation, followed by the formation of a second population of bigger amorphous particles that grow via addition of significantly smaller “monomers”, the nature of which remains as yet unknown (clusters and/or smaller units). The latter particles transform into the periodic ZIF...

This study aims to elucidate the aggregation and agglomeration behavior of TiO2 and ZrO2 nanoparticles during the nonaqueous synthesis. We found that zirconia nanoparticles immediately form spherical-like aggregates after nucleation with a homogeneous size of 200 nm, which can be related to the metastable state of the nuclei and the reduction of surface free energy. These aggregates further agglomerate, following a diffusion-limited colloid agglomeration mechanism that is additionally supported by the high fractal dimension of the resulting agglomerates. In contrast, TiO2 nanoparticles randomly orient and follow a reaction-limited colloid agglomeration mechanism that leads to a dense network of particles throughout the entire reaction volume. We performed in situ laser light transmission measurements and showed that particle formation starts earlier than previously reported. A complex population balance equation model was developed that is able to simulate particle aggregation as well as agglomeration, wh...

Intermediate filaments are a major structural element in the cytoskeleton of animal cells that mechanically integrate other cytoskeletal components and absorb externally applied stress. Their role is likely to be linked to their complex molecular architecture which is the product of a multi-step assembly pathway. Intermediate filaments form tetrameric subunits which assemble in the presence of monovalent salts to form unit length filaments that subsequently elongate by end-to-end annealing. The present work characterizes this complex assembly process using reconstituted vimentin intermediate filaments with monovalent salts as an assembly trigger. A multi-scale approach is used, comprising static light scattering, dynamic light scattering and quantitative scanning transmission electron microscopy (STEM) mass measurements. Light scattering reveals the radius of gyration (Rg), molecular weight (Mw) and diffusion coefficient (D) of the assembling filaments as a function of time and salt concentration (cS) for the given protein concentration of 0.07 g L−1. At low cS (10 mM KCl) no lateral or elongational growth is observed, whereas at cS = 50–200 mM, the hydrodynamic cross-sectional radius and the elongation rate increases with cS. Rgversus Mw plots suggest that the mass per unit length increases with increasing salt content, which is confirmed by STEM mass measurements. A kinetic model based on rate equations for a two step process is able to accurately describe the variation of mass, length and diffusion coefficient of the filaments with time and provides a consistent description of the elongation accelerated by increasing cS.

Dodecyl amine edge functionalized few-layer graphene oxide quantum dots were synthesized in good yields. The covalent functionalization was demonstrated with NMR and AFM-IR. The resulting structure and particle size was measured with AFM and HRTEM. The thermal stability of the compound was investigated and showed a stability of up to 220 °C. The modified graphene oxide quantum dots showed excellent solubility in various organic solvents, including ethers, methanol, toluene, n-hexane, heptane, xylene, dichloromethane and toluene. The stability of a resulting toluene solution was also proven by static light scattering measurements over several days. The excellent solubility gives the possibility of an efficient and fast spray application of the functionalized graphene oxide quantum dots to steel surfaces. Hence, the macroscopic friction behavior was investigated with a Thwing-Albert FP-2250 friction tester. A thin film of the dodecyl amine functionalized graphene oxide quantum dots on steel lowered the friction coefficient from 0.17 to 0.11 and revealed a significant corrosion inhibition effect.

Dilute solutions of sodium poly(styrene sulfonate) (NaPSS) in the presence of Al3+, Ca2+, and Ba2+ were analysed by means of isothermal titration calorimetry (ITC) in order to investigate the heat effect of bond formation between those cations and the anionic SO3− residues of NaPSS. The selection of the cations was guided by the solution behavior of the corresponding PSS salts from a preceding study [M. Hansch et al., J. Chem. Phys. 148(1), 014901 (2018)], where bonds between Ba2+ and anionic PSS showed an increasing solubility with decreasing temperature and Al3+ exhibited the inverse trend. Unlike to Al3+ and Ba2+, Ca2+ is expected to behave as a purely electrostatically interacting bivalent cation and was thus included in the present study. Results from ITC satisfactorily succeeded to explain the temperature-dependent solution behavior of the salts with Al3+ and Ba2+ and confirmed the non-specific behavior of Ca2+. Additional ITC experiments with salts of Ca2+ and Ba2+ and sodium poly(acrylate) complemented the results on PSS by data from a chemically different polyanion. Availability of these joint sets of polyanion-cation combinations not only offers the chance to identify common features and subtle differences in the solution behavior of polyelectrolytes in the presence of multi-valent cations but also points to a new class of responsive materials.Dilute solutions of sodium poly(styrene sulfonate) (NaPSS) in the presence of Al3+, Ca2+, and Ba2+ were analysed by means of isothermal titration calorimetry (ITC) in order to investigate the heat effect of bond formation between those cations and the anionic SO3− residues of NaPSS. The selection of the cations was guided by the solution behavior of the corresponding PSS salts from a preceding study [M. Hansch et al., J. Chem. Phys. 148(1), 014901 (2018)], where bonds between Ba2+ and anionic PSS showed an increasing solubility with decreasing temperature and Al3+ exhibited the inverse trend. Unlike to Al3+ and Ba2+, Ca2+ is expected to behave as a purely electrostatically interacting bivalent cation and was thus included in the present study. Results from ITC satisfactorily succeeded to explain the temperature-dependent solution behavior of the salts with Al3+ and Ba2+ and confirmed the non-specific behavior of Ca2+. Additional ITC experiments with salts of Ca2+ and Ba2+ and sodium poly(acrylate) complem...

The final step in the supramolecular buildup of eumelanin particles is investigated regarding the involved species and mechanism. Time-resolved in situ light scattering and scanning electron microscopy reveal an aggregation of particles with a narrow size distribution around 40 nm, previously only observed as substructures. These form larger particles with again very uniform size and diameters around 200 nm. Aggregation of each single particle takes only a few minutes to complete, whereas the entire process goes on for at least 3 h, partly due to the kinetics of the precursors. The individual particles also undergo an additional consolidation step toward their final form, which takes up to 24 h. Atomic force microscopy shows that the size before consolidation is around twice the size of the final particles, due to free space between the substructures. Light scattering also reveals that the aggregation is random with respect to where the particles attach, as the shape of aggregates changes from sphere to coil, before it returns to a spherical shape at the end. Application of enzyme mediated autodeposition finally shows the potential to stop the supramolecular buildup at each level, and therefore enables isolation of the respective eumelanin particles at will. This may enable the full potential for melanin materials in nanotechnology deriving from its unique (for biological polymers) properties like paramagnetism, electrical conductivity, and many more.

The early stages of silica polymerization in aqueous solution proceed according to a mechanism based on three steps: nucleation, particle growth, and agglomeration of the particles. Application of time-resolved static and dynamic light scattering as a powerful in situ technique in combination with spectrophotometric analysis of the monomer consumption based on the molybdenum blue method was carried out to further investigate this 3-step process. Experiments were carried out at four different initial silicic acid contents covering a range between 350 and 750 ppm in the presence of either 10 mM NaCl or 5 mM of a mixture of CaCl2 and MgCl2. The process in all cases was initiated with a drop of pH to 7. Addition of the salts made possible an analysis of the impact of an electrolyte on the process. Independent of the presence or absence of salt, particle growth in step two proceeded as a monomer-addition process without being interfered significantly by Ostwald-ripening. The growing particles were compact with...