Refolding of Denatured‐Reduced Lysozyme in the Presence of Native Templates

Abstract

Protein refolding is known to be dependent on various factors, some of the important ones are the concentration of the protein sample, temperature, pH and also the concentration of the denaturant. Protein folding and aggregation are competing reactions and the factors responsible for the partition between these two reactions are still not well understood. One factor that has not been studied is the effect of a folded protein on the refolding of denatured state/form(s) of the same protein. This study aims to determine the degree to which protein refolding is template-driven by studying the refolding of lysozyme (a model protein). The effects of adding a folded template (native state), on the renaturation yield of denatured states of lysozyme was ascertained by biological assays, high performance liquid chromatography (HPLC), and intrinsic fluorescence studies. We have observed that at a template concentration of 40 µg/mL and denatured-reduced protein concentration of 160 µg/mL that there is an approximate 3 % increase in refolding yield based on biological assays. Since protein refolding is such a critical cellular process and given that protein aggregation is at the root of numerous diseases, the gain from this study will provide useful information on the factors that govern protein refolding, which could be extended to minimize the process of protein aggregation.