Conformational Consequences for Compatible Osmolytes on Thermal Denaturation.

Compatible osmolytes are a broad class of small organic molecules employed by living systems to combat environmental stress by enhancing the native protein structure. The molecular features that make for a superior biopreservation remain elusive. Through the use of time-resolved and steady-state spectroscopic techniques, in combination with molecular simulation, insight into what makes one molecule a more effective compatible osmolyte can be gained.

Hydration Dynamics in Solutions of Cyclic Polyhydroxyl Osmolytes.

Simple sugars are remarkably effective at preserving protein and enzymatic structures against thermal and hydrostatic stress. Here, we investigate the hydrodynamic and biopreservative properties of three small cyclic molecules: glucose, -inositol, and methyl-α-d-glucopyranoside using circular dichroism spectroscopy and isothermal calorimetry. Using ultrafast fluorescence frequency upconversion spectroscopy, we measure the dynamical retardation of hydration dynamics in cosolute solutions.

Hydrophobic interactions of sucralose with protein structures.

Sucralose is a commonly employed artificial sweetener that appears to destabilize protein native structures. This is in direct contrast to the bio-preservative nature of its natural counterpart, sucrose, which enhances the stability of biomolecules against environmental stress. We have further explored the molecular interactions of sucralose as compared to sucrose to illuminate the origin of the differences in their bio-preservative efficacy.

Retardation of Bulk Water Dynamics by Disaccharide Osmolytes.

The bioprotective nature of disaccharides is hypothesized to derive from the modification of the hydrogen bonding network of water which protects biomolecules through lowered water activity at the protein interface. Using ultrafast fluorescence spectroscopy, we measured the relaxation of bulk water dynamics around the induced dipole moment of two fluorescent probes (Lucifer Yellow Ethylenediamine and Tryptophan). Our results indicate a reduction in bulk water reorganization rate of approximately 30%.

Sucralose Destabilization of Protein Structure.

Sucralose is a commonly employed artificial sweetener that behaves very differently than its natural disaccharide counterpart, sucrose, in terms of its interaction with biomolecules. The presence of sucralose in solution is found to destabilize the native structure of two model protein systems: the globular protein bovine serum albumin and an enzyme staphylococcal nuclease. The melting temperature of these proteins decreases as a linear function of sucralose concentration.