Metal-mediated self-assembly of protein superstructures: influence of secondary interactions on protein oligomerization and aggregation

We have previously demonstrated that non-self-associating protein building blocks can oligomerize to form discrete supramolecular assemblies under the control of metal coordination. We show here that secondary interactions (salt bridges and hydrogen bonds) can be critical in guiding the metal-induced self-assembly of proteins. Crystallographic and hydrodynamic measurements on appropriately engineered cytochrome cb562 variants pinpoint the importance of a single salt-bridging arginine side chain in determining whether the protein monomers form a discrete Zn-induced tetrameric complex or heterogeneous aggregates. The combined ability to direct PPIs through metal coordination and secondary interactions should provide the specificity required for the construction of complex protein superstructures and the selective control of cellular processes that involve protein-protein association reactions.     

Polymers for in vivo tuning of refractive properties in intraocular lenses

High-refractive-index polymers for intraocular lenses are presented whose refractive index can be tuned post-operatively in a non-invasive manner to enable focal length and aspheric corrections in vivo. The polymer materials contain covalently attached photochemically crosslinkable coumarin side groups, to cause a change in refractive index of Deltan > 0.02, which enables one to tune the focal length of a standard IOL by two diopters. This is enough to ensure that more than 80% of all cataract patients can be adjusted non-invasively to optimal vision after cataract surgery, thus providing a remedy to a major shortcoming of today's cataract surgery.     

Extractive Spectrophotometric Methods for Determination of Clarithromycin in Pharmaceutical Formulations Using Bromothymol Blue and Cresol Red

Two simple and sensitive extractive spectrophotometric methods have been described for the analysis of clarithromycin in pure form and in pharmaceutical formulations. The methods involved formation of yellow colored chloroform extractable ion‐association complexes of clarithromycin with bromothymol blue (BTB) and cresol red (CR) in buffered aqueous solution at pH 4. The extracted complexes showed maximum absorbance at 410 and 415 nm for BTB and CR, respectively. Beer's law is obeyed in the concentration ranges 0.1–20 μg mL^?1 and 2.0–20 μg mL^?1 of clarithromycin with molar absorptivity of 2.01 × 10⁴ and 4.378 × 10³ for BTB and CR, respectively. The composition ratio of the ion‐association complex was clarithromycin: BTB and CR = 1:1 as established by Job's method. The methods have been applied to the determination of drug in commercial formulations. The results of analysis were validated statistically and through recovery studies.     

Distinct Spacing Between Anionic Groups: An Essential Chemical Determinant for Achieving Thiophene‐Based Ligands to Distinguish β‐Amyloid or Tau Polymorphic Aggregates

The accumulation of protein aggregates is associated with many devastating neurodegenerative diseases and the existence of distinct aggregated morphotypes has been suggested to explain the heterogeneous phenotype reported for these diseases. Thus, the development of molecular probes able to distinguish such morphotypes is essential. We report an anionic tetrameric oligothiophene compound that can be utilized for spectral assignment of different morphotypes of β‐amyloid or tau aggregates present in transgenic mice at distinct ages. The ability of the ligand to spectrally distinguish between the aggregated morphotypes was reduced when the spacing between the anionic substituents along the conjugated thiophene backbone was altered, which verified that specific molecular interactions between the ligand and the protein aggregate are necessary to detect aggregate polymorphism. Our findings provide the structural and functional basis for the development of new fluorescent ligands that can distinguish between different morphotypes of protein aggregates.     

Extracting [Pd@Sn9]4– and [Rh@Pb9]4– Clusters from their Binary Alloys Using “Metal Scissors”

The search for novel ternary intermetallic compounds with specific structures is still a challenge. We found that the two-step synthesis giving a typical alloy in the first step followed by the reaction of this alloy with alkali metals is a promising route. The intermetallic compounds K₁₂Pd_0.47Sn₁₇ and K₄RhPb₉ were synthesized by high-temperature reactions of preformed Pd-Sn and Rh-Pb alloys with K acting as “metal scissors” and were characterized by means of single crystal and powder X-ray diffractometry. The salt-like ternary phases K₁₂Pd_0.47Sn₁₇ and K₄RhPb₉ contain novel endohedrally filled intermetalloid clusters [Pd@Sn₉]^4– and [Rh@Pb₉]^4–, respectively. The crystal packing of the products corresponds to filled variants of the binary Zintl phases K₁₂Sn₁₇ and K₄Pb₉, respectively. The crystal structure of K₁₂Pd_0.47Sn₁₇ can be regarded as a hierarchical replacement variant of the hexagonal Laves phase MgZn₂, with [Pd@Sn₉]^4–/[Sn₉]^4– and [Sn₄]^4– on Mg and Zn positions, respectively, whereas the packing of [Rh@Pb₉]^4– clusters in K₄RhPb₉ is hcp. K₁₂Pd_0.47Sn₁₇ was characterized by Raman spectroscopy. For the first time Raman modes typical for endohedrally filled [Pd@Sn₉]^4– clusters are observed and in accordance with quantum-chemical calculations. In addition Raman spectroscopy shows also the presence of filled Pd@Sn₉ clusters in a phase of nominal composition “Na₁₂Pd₂Sn₁₇”. The results are discussed with respect to the volume increase due the incorporation of transition metal atoms.     

Convolutions of multivariate phase-type distributions

This paper is concerned with multivariate phase-type distributions introduced by Assaf et al. (1984). We show that the sum of two independent bivariate vectors each with a bivariate phase-type distribution is again bivariate phase-type and that this is no longer true for higher dimensions. Further, we show that the distribution of the sum over different components of a vector with multivariate phase-type distribution is not necessarily multivariate phase-type either, if the dimension of the components is two or larger.     

Rheology of polyacrylate binders produced via catalytic chain transfer polymerization as an alternative to bitumen in road pavement materials

Catalytic chain transfer polymerization has been successfully used to produce a range of methyl acrylate (MA) and butyl acrylate (BA) synthetic polymers of specific, targeted molecular weights, with polydispersity index values in the range of 2–4.5. The rheological properties of a subgroup of these synthetic binders consisting of four MA homo-polymers and one MA–BA co-polymer were then determined by means of oscillatory testing using a dynamic shear rheometer (DSR). The rheological tests consisted of a combination of stress/strain amplitude and frequency sweeps using a standard 8 mm diameter parallel plate testing geometry. The rheological parameters of phase angle and complex, storage and loss moduli were then shifted to form master curves at a reference temperature of 25 °C and isochronal plots at 0.1, 1 and 10 Hz. The rheological properties of the synthetic polymers were also compared to those of standard road pavement bitumens. The results show that it is possible to produce a range of synthetic polyacrylates with different rheological responses by altering the reactant type, reactant concentration and polymerization conditions to match the rheological properties of road bitumens. All the polyacrylate binders showed a similar rheological profile with a unique viscoelastic response as represented by the phase angle master curves together with an upper limiting stiffness and intermediate temperature/frequency ‘plateau’ region as shown in the complex modulus master curves. The results of the rheological examination of the binders showed that the key material property that influenced the performance of the polyacrylates in these specific application tests was glass transition temperature rather than molecular weight. Over this range of investigated molecular weights, it is the ratio between the two polymers which determines the glass transition and as such determines the material properties. These findings suggest that such sustainably sourced polyacrylate binders may allow for a move from petrochemical feed stocks to be made and allow for targeted road pavement design based on local climates, offering improved mechanical robustness.     

Inhibition of the histone demethylase JMJD2E by 3-substituted pyridine 2,4-dicarboxylates

Based on structural analysis of the human 2-oxoglutarate (2OG) dependent JMJD2 histone N(ε)-methyl lysyl demethylase family, 3-substituted pyridine 2,4-dicarboxylic acids were identified as potential inhibitors with possible selectivity over other human 2OG oxygenases. Microwave-assisted palladium-catalysed cross coupling methodology was developed to install a diverse set of substituents on the sterically demanding C-3 position of a pyridine 2,4-dicarboxylate scaffold. The subsequently prepared di-acids were tested for in vitro inhibition of the histone demethylase JMJD2E and another human 2OG oxygenase, prolyl-hydroxylase domain isoform 2 (PHD2, EGLN1). A subset of substitution patterns yielded inhibitors with selectivity for JMJD2E over PHD2, demonstrating that structure-based inhibitor design can enable selective inhibition of histone demethylases over related human 2OG oxygenases.     

Molecular approaches towards mixed metal oxides and their behaviour in mixed oxide support Au catalysts for CO oxidation

We herein report a water-based sol-gel approach towards porous mixed Si/Ti oxides using co-precipitated glycol-modified precursors. By adjusting synthesis parameters such as the pH value and the Si/Ti ratio of the precursor, the morphology as well as the Si/Ti-composition of the resulting mixed oxide particles can be varied in a wide range. The behaviour of the mixed oxides as substrates for Au catalysts and the performance of the resulting catalysts in the CO oxidation reaction was investigated and compared to catalysts supported on mesoporous anatase and rutile synthesized analogously. For comparable Au particle sizes and Au loadings, the composition of the mixed oxide support was found to significantly affect the reactivity and reaction behaviour, with mixed oxide supported Au catalysts synthesized at pH=5 or 10 and with a Si/Ti-ratio of 1:19 and 1:34 exhibiting the maximum activity. In contrast to the enhanced activity, the mixed oxide supports do not lead to a significant improvement in deactivation behaviour and catalyst stability.