Coomassie Brilliant Blue Induces Coiled-Coil Aggregation in Lysozyme at pH 7.4 by Hydrophobic and Electrostatic Forces

考马斯亮蓝在 pH 7.4 时通过疏水作用和静电作用诱导溶菌酶发生卷曲螺旋聚集

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Abstract

Several neurodegenerative diseases are associated with the deposition of amyloid fibrils. Although these diseases are irreversible, knowing the aggregation mechanism is useful in developing drugs that can arrest or decrease the aggregation rate. In this study, we are interested in investigating the effect of Coomassie brilliant blue (CBB G-250) on the aggregation of hen egg white lysozyme (HEWL) at pH 7.4. Various biophysical techniques have been used, such as turbidity, Rayleigh light scattering (RLS) kinetics, far-UV circular dichroism (CD), field emission scanning electron microscopy (FESEM), and transmission electron microscopy (TEM) imaging. The turbidity data indicated that CBB (≥0.1 mM) induces aggregation in HEWL at pH 7.4. The aggregation kinetics caused by CBB are quick without a lag phase and are dependent on the CBB concentration. The far-UV CD data revealed that the CBB-induced aggregated samples had lost their CD signals without exhibiting a shift in the spectrum position. Sodium chloride and ammonium sulfate show little effect on the CBB-induced aggregates, but alcohol such as methanol, ethanol, and 2-propanol could reverse the aggregation. Overall, this study aims to better understand the mechanism underlying CBB-induced aggregation and keep in mind that CBB employed in laboratories can alter the protein structure. We report the aggregation of a natural protein due to coiled-coil formation induced by a dye at physiological pH and temperature conditions. This finding has high value because several dyes are used for diagnostic and therapeutic purposes, and coiled-coil formation is closely related to infection mechanisms and nanoparticle-based drug deliveries.

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