Analysis of sugar bioprotective mechanisms on the thermal denaturation of lysozyme from Raman scattering and differential scanning calorimetry investigations.
Sugar-induced thermostabilization of lysozyme was analyzed by Raman scattering and modulated differential scanning calorimetry investigations, for three disaccharides (maltose, sucrose, and trehalose) characterized by the same chemical formula (C(12)H(22)O(11)). This study shows that trehalose is the most effective in stabilizing the folded secondary structure of the protein. The influence of sugars on the mechanism of thermal denaturation was carefully investigated by Raman scattering experiments carried out both in the low-frequency range and in the amide I band region. It was determined that the thermal stability of the hydrogen-bond network of water, highly dependent on the presence of sugars, contributes to the stabilization of the native tertiary structure and inhibits the first stage of denaturation, that is, the transformation of the tertiary structure into a highly flexible state with intact secondary structure. It was found that trehalose exhibits exceptional capabilities to distort the tetra-bonded hydrogen-bond network of water and to strengthen intermolecular O-H interactions responsible for the stability of the tertiary structure. Trehalose was also observed to be the best stabilizer of the folded secondary structure, in the transient tertiary structure, leading to a high-temperature shift of the unfolding process (the second stage of denaturation). This was interpreted from the consideration that the transient tertiary structure is less flexible and inhibits the solvent accessibility around the hydrophobic groups of lysozyme.
Hédoux
A
A
Laboratoire de Dynamique et Structure des Matériaux Moléculaires, UMR CNRS 8024, Université de Lille 1, UFR de Physique, Bat. P5, 59 655 Villeneuve d'Ascq Cedex, France. alain.hedoux@univ-lille1.fr
Willart
J-F
JF
Ionov
R
R
Affouard
F
F
Guinet
Y
Y
Paccou
L
L
Lerbret
A
A
Descamps
M
M
D000577:Amides / Q000737:chemistry; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002241:Carbohydrates / Q000032:analysis / Q000737:chemistry*; D004795:Enzyme Stability; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D009113:Muramidase / Q000032:analysis / Q000737:chemistry*; D011489:Protein Denaturation; D012997:Solvents / Q000737:chemistry; D013059:Spectrum Analysis, Raman / Q000379:methods; D013696:Temperature; D014199:Trehalose / Q000032:analysis / Q000737:chemistry; D014867:Water / Q000737:chemistry
D000577:Amides; D002241:Carbohydrates; D012997:Solvents; D014867:Water; D014199:Trehalose; D009113:Muramidase