Exploring the role of urea in preventing aggregation in alpha Lactalbumin protein using biophysical approaches

Protein aggregation is a common characteristic of numerous neurodegenerative diseases and presents challenges in biotechnology and pharmaceutical formulations. Urea, a well-known denaturant, has been widely studied for its role in protein folding and unfolding. However, its potential role in preventing protein aggregation and mitigating macromolecular crowding effects remains less explored. In this study, we investigate the molecular mechanisms by which lower concentration of urea influences protein aggregation in crowded environments. Using a combination of spectroscopic techniques, microscopic, and aggregation assays, we demonstrate that urea modulates protein-solvent interactions, reduces intermolecular contacts that drive aggregation, and alleviates excluded volume effects in crowded solutions. Our findings suggest that urea acts not only as a chemical chaperone but also as a tunable modulator of protein solubility in physiologically relevant conditions. These insights provide a deeper understanding of urea's multifaceted role in protein chemistry and may have implications for therapeutic strategies targeting protein misfolding diseases.