American Chemical Society, Biochemistry, 19(54), p. 2957-2966, 2015
DOI: 10.1021/acs.biochem.5b00116
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We examined the effects of water-soluble polymers of various degrees of hydrophobicity on folding and aggregation of proteins. The polymers we chose were polyethylene glycol (PEG) and UCON (1:1 copolymer of ethylene glycol and propylene glycol). Presence of additional methyl groups in UCON makes it more hydrophobic than PEG. Our earlier analysis revealed that similarly sized PEG and UCON produced different changes in the solvent properties of water in their solutions and induced morphologically different α-synuclein aggregates (Ferreira et al. Role of solvent properties of aqueous media in macromolecular crowding effects. Journal of Biomolecular Structure and Dynamics. In press, (1)). In order to better understand molecular mechanisms defining behavior of proteins in crowded environment, we tested the effects of these polymers on secondary and tertiary structure and aromatic residue solvent accessibility of 10 proteins (five folded proteins, two hybrid proteins; i.e., protein containing ordered and disordered domains, and three intrinsically disordered proteins, IDPs) and on aggregation kinetics of insulin and α-synuclein. We found that effects of both polymers on secondary and tertiary structures of folded and hybrid proteins were rather limited with slight unfolding observed in some cases. Solvent accessibility of aromatic residues was significantly increased for the majority of the studied proteins in the presence of UCON but not PEG. PEG also accelerated protein aggregation into amyloid fibrils, whereas UCON promoted aggregation to amyloid oligomers instead. These results indicate that even a relatively small change in polymer structure leads to significant change in the effect of this polymer on protein folding and aggregation. This is an indication that protein folding and especially aggregation are highly sensitive to presence of other macromolecules, and excluded volume effect is insufficient to describe their effect.