Protein-Sugar-Water Systems Far From Thermodynamic Ideality

BSA Aggregation in Trehalose-Water Systems

Introduction Recent studies [1] on ternary protein-trehalose-water samples have shown that the protein thermal denaturation temperature is linearly correlated with the glass transition temperature of the system, despite the quite large temperature difference between the two processes. In such studies it is stated that the collective, long range properties of the matrix that regulate the glass transition are strictly correlated with local features on which the denaturation of the protein depends. In order to ascertain whether an analogous correlation exists between the effects of trehalose on the protein’s aggregation process and the glass transition temperature of the system, we performed L…

Thermal aggregation of BSA in trehalose or saccharose solutions

In Vitro–In Vivo Fluctuation Spectroscopies

Fluorescence correlation spectroscopy (FCS) was first developed for biophysical studies in analogy with photon scattering correlation spectroscopy. Although it is mainly devoted to the study of freely diffusing particles, FCS is actually able to discern between different kinds of motions, such as diffusion, anomalous diffusion, or drift motions. The frontier application of FCS nowadays is in medical studies both within cells and on the cell membranes, and in the investigation of single molecules in solid matrices. In this field, FCS originated also image correlation spectroscopy methods. The whole field can be unified under the name of fluorescence fluctuation spectroscopy (FFS). We present…

SAXS Study on Myoglobin Embedded in Amorphous Saccharide Matrices

We report on Small Angle X-ray Scattering (SAXS) measurements performed on samples of carboxy-myoglobin and met-myoglobin embedded in low hydrated matrices of four different saccharides (trehalose, sucrose, maltose and lactose). Results confirm the already reported occurrence of inhomogeneities, which are not peculiar of trehalose samples, but appear also in maltose and lactose, and in some cases also sucrose, being dependent on the sample hydration and on the presence of sodium dithionite. This behaviour confirms our previous interpretation about the nature of the inhomogeneities, and prompt it as a possible general behaviour for highly concentrated sugar matrices.

SAXS and FTIR study on MbCO-Saccharide amorphous systems

Thermal Aggregation of Bovine Serum Albumin in Trehalose and Sucrose Aqueous Solutions

We report results of static and dynamic light scattering measurements performed on bovine serum albumin (BSA) in saccharide (trehalose and sucrose) solutions. Our aim is to study the effects of the two disaccharides on the first steps of thermal aggregation of BSA in aqueous solutions at two protein concentrations (1 and 30 mg/mL) at increasing sugar/water ratio. Results show that sugars modify early stages of aggregation mainly by perturbing the thermodynamic behavior of the solvent (i.e., general solvent effects) without involving direct, specific sugar-protein interactions. This agrees with current hypotheses on sugar action in protein solutions. (1-3) The linear correlation detected bet…