In this work the effect of betaine on the structure and aggregation of the GST-GFP fluorescent fusion protein was studied by different complementary techniques, including electron microscopy, dynamic light scattering, circular dichroism, and FTIR spectroscopy. Although osmolytes are known to be protein stabilizers in vivo, the effect of betaine on the structure and aggregation of our model protein was found to be strictly concentration dependent. We demonstrated that, by changing betaine concentration, it was possible to tune the formation of protein soluble assemblies and insoluble aggregates, as well as to disaggregate preformed aggregates. In particular, at a critical concentration of betaine between 5 and 7.5 mM, the protein precipitated into macroscopic prefibrillar structures, rich in intermolecular P-sheets, which were found to bind thioflavine T and to be inaccessible to protease. Instead, at higher betaine concentration (1020 mM) the misfolded protein lost its fluorescence, but formed soluble assemblies with hydrodynamic radius of about 16 nm. These structures displayed a reduced propensity to further aggregate under thermal treatment. In addition, betaine at this high concentration was also found to disrupt large preformed aggregates, obtained under different conditions, into protein soluble assemblies. It is the first time that a disag-gregation process has been described for a chemical chaperone. A mechanism for the betaine concentration-dependent effect on protein misfolding, aggregation, and disaggregation is proposed and its possible physiological implications are discussed. © 2008 Wiley-Liss, Inc.
Natalello, A., Liu, J., Ami, D., Doglia, S.M., & De Marco, A. (2009). The osmolyte betaine promotes protein misfolding and disruption of protein aggregates. PROTEINS, 75(2), 509-517.
|Citazione:||Natalello, A., Liu, J., Ami, D., Doglia, S.M., & De Marco, A. (2009). The osmolyte betaine promotes protein misfolding and disruption of protein aggregates. PROTEINS, 75(2), 509-517.|
|Tipo:||Articolo in rivista - Articolo scientifico|
|Carattere della pubblicazione:||Scientifica|
|Titolo:||The osmolyte betaine promotes protein misfolding and disruption of protein aggregates|
|Autori:||Natalello, A; Liu, J; Ami, D; Doglia, SM; De Marco, A|
|Data di pubblicazione:||gen-2009|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1002/prot.22266|
|Appare nelle tipologie:||01 - Articolo su rivista|