Influence of crowding agents on the dynamics of a multidomain protein in its denatured state: a solvation approach

It is now well appreciated that the crowded intracellular environment significantly modulates an array of physiological processes including protein folding–unfolding, aggregation, and dynamics to name a few. In this work we have studied the dynamics of domain I of the protein human serum albumin (HSA) in its urea-induced denatured states, in the presence of a series of commonly used macromolecular crowding agents. HSA was labeled at Cys-34 (a free cysteine) in domain I with the fluorophore 6-bromoacetyl-2-dimethylaminonaphthalene (BADAN) to act as a solvation probe. In partially denatured states (2–6 M urea), lower crowder concentrations (~