Trehalose Restrains the Fibril Load towards α-Lactalbumin Aggregation and Halts Fibrillation in a Concentration-Dependent Manner

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.

Understanding the Modulation of α-Synuclein Fibrillation by N-Acetyl Aspartate: A Brain Metabolite

“…Osmolytes refer to small organic molecules that are released by cells in response to severe stress conditions . Numerous studies have shown that polyols osmolytes have a stabilizing effect on α-Syn monomers and inhibit the α-Syn aggregation. ,, However, the impact of amino acid osmolytes on α-Syn aggregation has not yet been elucidated yet. NAA is an amino acid osmolyte that is mainly present in the brain in high concentrations of up to 10 mM .…”

“…ThT, also known as thioflavin T, is a fluorescent dye commonly employed to monitor protein aggregation, owing to its ability to selectively bind to amyloid fibrils, which are protein aggregates commonly associated with neurodegenerative diseases . ThT was dissolved in water and filtered through a 0.22 μM syringe filter, and its concentration was determined by taking the absorbance at 412 nm using ε 412 = 24,420 M –1 cm . The aliquots withdrawn at specific time intervals from the incubated samples were diluted to 15 μM and incubated with 20 μM ThT for 15 min in the dark …”

“…33 ThT was dissolved in water and filtered through a 0.22 μM syringe filter, and its concentration was determined by taking the absorbance at 412 nm using ε 412 = 24,420 M −1 cm. 34 The aliquots withdrawn at specific time intervals from the incubated samples were diluted to 15 μM and incubated with 20 μM ThT for 15 min in the dark. 35 Three independent ThT fluorescence measurements of the sample were taken by a Jasco FP-8200 fluorescence spectrometer with an excitation wavelength of 440 nm and the emission wavelength in the range of 450−600 nm.…”

Impact of Single Amino Acid Substitutions in Parkinsonism-Associated Deglycase-PARK7 and Their Association with Parkinson’s Disease

“…The solubility of a protein influences its function to a great extent. Diseases such as Parkinson's disease, amyloidosis, and Alzheimer's disease are caused by the aggregation of insoluble parts of the proteins [30][31][32][33][34]. To predict the solubility of PARK7, we calculated the variants' solubility using SODA (Solubility based on Disorder and Aggregation).…”