Inhibition of lysozyme fibrillogenesis by hydroxytyrosol and dopamine: An Atomic Force Microscopy study

Abstract

Protein aggregation underlies many human diseases characterized by the deposition of normally soluble proteins in both fibrillar and amorphous aggregates. Here, Atomic Force Microscopy (AFM) has been applied to investigate the ability to inhibit hen egg white lysozyme (HEWL) fibrillogenesis by hydroxytyrosol (HT), one of the main phenolic components of olive oil. In this framework, HEWL is a useful and well-studied model protein whose amyloid-like fibril formation can be induced under experimental conditions where HT is more stable. HEWL fibrils, obtained at pH 1.6 and at 65 °C, exhibited a height of about 3 nm and a fibril length on average of about 3 μm. The presence of HT reduced the HEWL fibril number and length with respect to the control sample. Interestingly, also dopamine, a compound with a chemical structure similar to HT, decreased both the fibril number and the fibril length. AFM experimental data were supported by Thioflavin T assay and Fourier transform infrared spectroscopy. Our results show that HT is an effective inhibitor of HEWL aggregation, thus suggesting possible future applications of this natural compound for potential prevention or treatment of amyloid diseases, or as a lead molecular structure for the design of improved modulators.

Keywords: Amyloid; Atomic Force Microscopy; Dopamine; Hen egg white lysozyme; Hydroxytyrosol; Protein aggregation.