Thermal denaturation of beta-lactoglobulin and stabilization mechanism by trehalose analyzed from Raman spectroscopy investigations.
The thermal denaturation process of beta-lactoglobulin has been analyzed in the 20-100 degrees C temperature range by Raman spectroscopy experiments simultaneously performed in the region of amide modes (800-1800 cm(-1)) and in the low-frequency range (10-350 cm(-1)). The analysis of amide modes reveals a two-step thermal denaturation process in the investigated temperature range. The first step corresponds to the dissociation of dimers associated with an increase of flexibility of the tertiary structure. In the second step, large conformational changes are detected in the secondary structure and described as a loss of alpha-helix structures and a concomitant formation of beta-sheets. Raman investigations in the low-frequency range provide important information on the origin of the denaturation process through the analysis of the solvent dynamics and its coupling with that of the protein. The softening of the tetrahedral structure of water induces the dissociation of dimers and makes the tertiary structure softer, leading to the water penetration in the protein interior. The methodology based on Raman investigations of amide modes and in the low-frequency region was used to analyze the mechanism of beta-lactoglobulin thermostabilization by trehalose. The main effect of trehalose is determined to be related to its capabilities to distort the tetrahedral organization of water molecules.
Seo
Jeong-Ah
JA
Unité Matériaux Et Transformations, UMR CNRS 8207, Université de Lille 1, 59655 Villeneuve d Ascq Cédex, France.
Hédoux
Alain
A
Guinet
Yannick
Y
Paccou
Laurent
L
Affouard
Frédéric
F
Lerbret
Adrien
A
Descamps
Marc
M
D007782:Lactoglobulins / Q000737:chemistry*; D011489:Protein Denaturation; D055550:Protein Stability; D017433:Protein Structure, Secondary; D013059:Spectrum Analysis, Raman; D013696:Temperature; D014199:Trehalose / Q000737:chemistry*; D014867:Water / Q000737:chemistry
D007782:Lactoglobulins; D014867:Water; D014199:Trehalose