UWEC CERCA 2025

Biomolecular condensates (BMCs) are naturally occurring membraneless organelles formed through liquid-liquid phase separation (LLPS). They play significant roles in various cellular processes, including signal transduction, gene expression, and stress response. The thermodynamics of condensate formation involve a complex interplay between entropy and enthalpy. The loss in entropy due to ordered assembly formation inside the liquid-like condensate is compensated by the increase in intermolecular interaction enthalpy. The main factors that promote LLPS include changes in biomolecule concentration and intermolecular interactions. The LLPS process is sensitive to pH, temperature, and ionic strength. LLPS of intrinsically disordered proteins (IDPs) and unstructured domains/regions of proteins (IDPRs) are well documented. Recent studies suggest that globular proteins also form crowder-induced biomolecular condensates. However, the precise role of molecular crowding in LLPS-driven biomolecular condensate formation remains understudied, especially for globular proteins. A thorough understanding of the molecular mechanism of protein condensate formation and the impact of phase separation on enzymatic reactions is crucial to addressing issues related to cellular physiology. To investigate the molecular mechanism of condensate formation in crowded environments, globular proteins such as bovine serum albumin and prolyl-tRNA synthetase are being used alongside synthetic crowders like polyethylene glycols of variable sizes. Additionally, variable salt concentrations are employed to understand the effects of multivalent interactions in BMC formation. We will present the preliminary results of different globular protein samples with and without polyethylene glycol (PEG) as crowding agents at high and low salt concentrations.