title: A desolvation model for trifluoroethanol-induced aggregation of enhanced green fluorescent protein.
abstract: Studies of amyloid disease-associated proteins in aqueous solutions containing 2,2,2-trifluoroethanol (TFE) have shown that the formation of structural intermediates is often correlated with enhanced protein aggregation. Here, enhanced green fluorescent protein (EGFP) is used as a model protein system to investigate the causal relationship between TFE-induced structural transitions and aggregation. Using circular dichroism spectroscopy, light scattering measurements, and transmission electron microscopy imaging, we demonstrate that population of a partially α-helical, monomeric intermediate is roughly correlated with the growth of β-sheet-rich, flexible fibrils for acid-denatured EGFP. By fitting our circular dichroism data to a model in which TFE-water mixtures are assumed to be ideal solutions, we show that increasing entropic costs of protein solvation in TFE-water mixtures may both cause the population of the intermediate state and increase aggregate production. Tertiary structure and electrostatic repulsion also impede aggregation. We conclude that initiation of EGFP aggregation in TFE likely involves overcoming of multiple protective factors, rather than stabilization of aggregation-prone structural elements.
authors.affiliation: School of Applied and Engineering Physics, Cornell University, Ithaca, New York, USA.
authors.forename: Valerie L
authors.initials: VL
authors.lastname: Anderson
authors.forename: Watt W
authors.initials: WW
authors.lastname: Webb
mesh_terms: D000818:Animals; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D037402:Hydrozoa; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012997:Solvents / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D014270:Trifluoroethanol / Q000737:chemistry*
chemical_list: D012997:Solvents; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D014270:Trifluoroethanol