Abstract
Protein aggregation and misfolding are some of the most challenging obstacles, customarily studied for their association with amyloid pathologies. The mechanism of amyloid fibrillation development is a dynamic phenomenon involving various factors such as the intrinsic properties of protein and the physical and chemical environmental conditions. The purpose of this study was to see the thermal aggregation profile of alpha-lactalbumin (α-LA) and to delineate the effect of trehalose on its aggregation profile. α-LA was subjected to thermal aggregation at high concentrations. UV-Vis spectroscopy, a turbidity assay, intrinsic fluorescence, Rayleigh scattering and a thioflavin T (ThT) assay explained the steady outcomes that 1 M trehalose repressed α-LA aggregation in the most effective way followed by 0.75 M and 0.5 M and to a significantly lesser degree by 0.25 M. Multi spectroscopic obser Sania Bashir ations were further entrenched by microscopy. Transmission electron microscopy confirmed that in the presence of its higher concentration, trehalose hinders fibril development in α-LA. In vitro studies were further validated by in silico studies. Molecular docking analysis indicated that trehalose occupied the binding pocket cavity of α-LA and offered several significant interactions, including H-bonds with important residues. This study provides a platform for trehalose in the therapeutic management of protein aggregation-related diseases.
Highlights
Several neurodegenerative diseases leading to amyloid fibrils and plaques are unambiguously associated with the intracellular aggregation of proteins [1,2]
(1 + e(−( T − Tagg)/b) where A is the absorbance at any temperature, T, Ao is the absorbance of the initial baseline, Amax is the absorbance of the final plateau line, b is the constant independent of T at a given wavelength, and osmolyte concentration and Aagg is the temperature at 50% of the maximum absorbance occurrence
UV-Vis spectroscopy, a Thioflavin T (ThT) binding assay, intrinsic fluorescence, Rayleigh scattering and turbidity assay measurements showed that the presence of trehalose inhibited α-LA
Summary
Several neurodegenerative diseases leading to amyloid fibrils and plaques are unambiguously associated with the intracellular aggregation of proteins [1,2]. Notwithstanding, this fibrillation process experiences several events including halfway misfolding followed by the development of oligomers, protofibrils and long-run fibrils. Alpha-lactalbumin (α-LA), a Ca2+ metallo binding [3] milk protein, is a constituent of whey proteins. Whey proteins, including α-LA, are exposed to heat during the process of pasteurization. Heating extensively leads to the aggregation and gel development of proteins with both favorable and non-favorable effects depending upon the objective of utilizing protein ingredients [4]. The choice of an osmolyte, which potentially acts as a molecular aggregation blocker and contributes
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