Antiamyloid Activity of Functionalized Cerium Oxide Nanoparticle on Lysozyme Fibrillation: Spectroscopic and Microscopic Investigation

Abstract

Accumulation of amyloid fibrils of proteins in different organs is a pathological indication for various neurogenerative disorders. Recently nanotechnology is at the centre of interest to tackle with those disorders. In this study we have chosen hen egg white lysozyme as our model protein which is responsible for hereditary systemic Amyloidosis disease. Here we have reported effect of Cerium oxide nanoparticle (CeONP), folic acid functionalized CeONP (FA-CeONP) and polyacrylic acid functionalized CeONP (PAA-CeONP) (all of them having size ~18 nm) on fibrillation of hen egg white lysozyme (Lyz). The amyloid growth is monitored by Thioflavin T (ThT), Congo Red (CR), 8-anilinonapthalene-1-sulfonic acid (ANS) assay, time resolved fluorescence anisotropy, intrinsic fluorescence, circular dichroism, atomic force microscopy and fluorescence microscopy study. Results demonstrate that all nanoparticle (NP) shows antiamyloid activity by broadening lag phase and suppressing growth phase of Lyz amyloid growth, but PAA-CeONP shows maximum inhibitory effect. These bare and functionalized CeONP shows dose dependent antiamyloid activity. We have also demonstrated that such inhibition of amyloid formation is associated with reduction of s-sheet content of protein. The inhibition efficiency is quantified by half maximal inhibition concentration IC50 values. A mechanistic pathway is explained on the basis of electrostatic interaction between NP and Lyz by ζ- potential measurement. This interaction is also analyzed theoretically by docking analysis. Thus our study provides a basis for use of CeONP as a promising candidate in therapeutic application in amyloid related disorders.