title: Towards a molecular level understanding of protein stabilization: the interaction between lysozyme and sorbitol.
abstract: The paper is investigating the mechanism of stabilization of proteins by polyols at the molecular level. It is addressing the interactions of sorbitol, a polyol commonly used as a protein stabilizing agent, with hen egg white lysozyme, a well studied protein. Differential scanning calorimetry shows an increase in denaturation temperature of lysozyme upon addition of sorbitol at a concentration of 250 mM and above. Increasing sorbitol concentration also caused an increase in signal intensity of the CD spectrum of lysozyme in the wavelength region of 280-300 nm. Two-dimensional nuclear magnetic resonance spectroscopy was used to examine interactions between lysozyme and sorbitol. Most significant changes are manifest in the anomalous relaxation properties of Ala and Thr methyl groups indicating modifications of local motions and possibly compression of the entire structure. This is further corroborated by new intra-protein nuclear Overhauser effects in the presence of sorbitol. There is also evidence that water is displaced from the enzyme surface close to Ile-88 upon addition of sorbitol. In combination these results reveal a complex interplay of different interactions. Comparison to NMR-spectra of lysozyme with a bound inhibitor (tri-N-acetyl-glucosamine) shows that the interaction with sorbitol affects spatially disparate regions of the protein.
authors.affiliation: MR-Center, SINTEF-UNIMED, Trondheim, Norway.
authors.forename: H
authors.initials: H
authors.lastname: Wimmer
authors.forename: M
authors.initials: M
authors.lastname: Olsson
authors.forename: M T
authors.initials: MT
authors.lastname: Petersen
authors.forename: R
authors.initials: R
authors.lastname: Hatti-Kaul
authors.forename: S B
authors.initials: SB
authors.lastname: Peterson
authors.forename: N
authors.initials: N
authors.lastname: Müller
mesh_terms: D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D004795:Enzyme Stability; D009682:Magnetic Resonance Spectroscopy; D009113:Muramidase / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D013012:Sorbitol / Q000737:chemistry*; D013696:Temperature; D013816:Thermodynamics
chemical_list: D011506:Proteins; D012997:Solvents; D013012:Sorbitol; D009113:Muramidase