Dynamics of C-phycocyanin in various deuterated trehalose/water environments measured by quasielastic and elastic neutron scattering.
The molecular understanding of protein stabilization by the disaccharide trehalose in extreme temperature or hydration conditions is still debated. In the present study, we investigated the role of trehalose on the dynamics of the protein C-phycocyanin (C-PC) by neutron scattering. To single out the motions of C-PC hydrogen (H) atoms in various trehalose/water environments, measurements were performed in deuterated trehalose and heavy water (D2O). We report that trehalose decreases the internal C-PC dynamics, as shown by a reduced diffusion coefficient of protein H atoms. By fitting the Elastic Incoherent Structure Factor--which gives access to the "geometry" of the internal proton motions--with the model of diffusion inside a sphere, we found that the presence of trehalose induces a significantly higher proportion of immobile C-PC hydrogens. We investigated, by elastic neutron scattering, the mean square displacements (MSDs) of deuterated trehalose/D2O-embedded C-PC as a function of temperature in the range of 40-318 K. Between 40 and approximately 225 K, harmonic MSDs of C-PC are slightly smaller in samples containing trehalose. Above a transition temperature of approximately 225 K, we observed anharmonic motions in all trehalose/water-coated C-PC samples. In the hydrated samples, MSDs are not significantly changed by addition of 15% trehalose but are slightly reduced by 30% trehalose. In opposition, no dynamical transition was detected in dry trehalose-embedded C-PC, whose hydrogen motions remain harmonic up to 318 K. These results suggest that a role of trehalose would be to stabilize proteins by inhibiting some fluctuations at the origin of protein unfolding and denaturation.
Köper
Ingo
I
Laboratoire Léon-Brillouin UMR 12 CEA/CNRS, CEA/Saclay, 91191, Gif-sur-Yvette Cedex, France.
Combet
Sophie
S
Petry
Winfried
W
Bellissent-Funel
Marie-Claire
MC
D003198:Computer Simulation; D003903:Deuterium / Q000737:chemistry*; D004548:Elasticity; D008956:Models, Chemical; D008958:Models, Molecular; D033363:Neutron Diffraction / Q000379:methods*; D010798:Phycocyanin / Q000737:chemistry* / Q000648:ultrastructure*; D014199:Trehalose / Q000737:chemistry*; D014867:Water / Q000737:chemistry*
D014867:Water; D010798:Phycocyanin; D003903:Deuterium; D014199:Trehalose