Understanding the factors that give rise to tau aggregation and reactive oxygen species (ROS) is the key aspect in Alzheimer’s disease pathogenesis. Microtubule (MT) binding repeats of tau protein were suggested to play a critical role in tau aggregation. Here, we show that the interaction of Cu²⁺ with full-length MT binding repeats R1–R4 leads to the aggregation, and a Cys-based redox chemistry is critically involved in tau aggregation leading to disulfide-bridge dimerization of R2 and R3 and further aggregation into a fibrillar structure. Notably, ascorbate and glutathione, the most abundant antioxidants in neurons, cannot prevent the effect of Cu²⁺ on R2 and R3 aggregation. Detailed ESI-MS and NMR experiments demonstrate the interaction of Cu²⁺ with MT binding repeats. We show that redox activity of copper increases when bound to the MT repeats leading to ROS formation, which significantly contribute to cellular damage and neuron death. Results presented here provide new insights into the molecular mechanism of tau aggregation and ROS formation and suggest a new target domain for tau aggregation inhibitors.

Chlorinated paraffins (CPs) can be mixtures of nearly a half million possible isomers. Despite extensive use of CPs, their isomer composition and effects on the environment remain poorly understood. Here, we reveal the isomeric distributions of nine CP mixtures with single chain lengths (C14/15) and varying chlorination degrees. The molar distributions of CnH2n+2-mClm in each mixture was determined using high-resolution mass spectrometry (MS). Next, the mixtures were analyzed by applying both one-dimensional (1D) 1H, 13C and two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy. Due to substantially overlapping signals in the experimental NMR spectra, direct assignment of individual isomers was not possible. As such, a new NMR spectral matching approach was developed that used massive NMR databases predicted by a neural network algorithm to provide the top 100 most likely structural matches. The top 100 isomers appear to be an adequate representation of the overall mixture. Their modelled physicochemical and toxicity parameters agree with previous experimental results. Chlorines are not evenly distributed in any of the CP mixtures and show general preference at the third carbon. The approach described here can play a key role in understanding of complex isomeric mixtures such as CPs that cannot be resolved by MS alone.

Dissolved organic matter (DOM) chemistry and the potential for organic matter (OM) to self-associate with other OM components are important aspects of understanding the mechanisms of DOM sorption to clay surfaces. To investigate this further, we sorbed DOM isolated from peat humic acid onto either kaolinite, montmorillonite and gibbsite via ten sequential batch equilibration sorption experiments. Dissolved organic carbon (DOC) sorption to all minerals increased consistently, suggesting that sorption occurred via mineral-OM interactions at the beginning of the experiment. After six successive DOM loadings, the concentration of DOC sorbed by kaolinite and gibbsite began to plateau, likely due to the saturation of mineral surface sorption sites. Solution-state nuclear magnetic resonance (NMR) analysis of unbound DOM showed that kaolinite and montmorillonite sorbed aliphatic, protein and lignin components initially and primarily aliphatic and aromatic constituents in later sorption experiments, whereas gibbsite sorbed mostly aliphatic compounds during all DOM loadings. Analysis of the organo-clay complexes using ¹H high resolution–magic angle spinning (HR-MAS) NMR confirmed the preferential sorption of aromatic and aliphatic components to all three minerals. Overall, these results suggest that OM-OM interactions may be important mechanisms of DOM sorption to clay mineral surfaces.