Preferential hydration of lysozyme in water/glycerol mixtures: a small-angle neutron scattering study.
In solution small-angle neutron scattering has been used to study the solvation properties of lysozyme dissolved in water/glycerol mixtures. To detect the characteristics of the protein-solvent interface, 35 different experimental conditions (i.e., protein concentration, water/glycerol fraction in the solvent, content of deuterated compounds) have been considered and a suitable software has been developed to fit simultaneously the whole set of scattering data. The average composition of the solvent in the close vicinity of the protein surface at each experimental condition has been derived. In all the investigated conditions, glycerol resulted especially excluded from the protein surface, confirming that lysozyme is preferentially hydrated. By considering a thermodynamic hydration model based on an equilibrium exchange between water and glycerol from the solvation layer to the bulk, the preferential binding coefficient and the excess solvation number have been estimated. Results were compared with data previously derived for ribonuclease A in the same mixed solvent: even if the investigated solvent compositions were very different, the agreement between data is noticeable, suggesting that a unique mechanism presides over the preferential hydration process. Moreover, the curve describing the excess solvation number as a function of the solvent composition shows the occurrence of a region of maximal hydration, which probably accounts for the changes in protein stability detected in the presence of cosolvents.
Sinibaldi
Raffaele
R
Dipartimento di Scienze Applicate ai Sistemi Complessi, Università Politecnica delle Marche, Ancona 60131, Italy.
Ortore
Maria Grazia
MG
Spinozzi
Francesco
F
Carsughi
Flavio
F
Frielinghaus
Henrich
H
Cinelli
Stefania
S
Onori
Giuseppe
G
Mariani
Paolo
P
D000818:Animals; D002645:Chickens; D004529:Egg White / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry*; D008956:Models, Chemical; D015233:Models, Statistical; D009113:Muramidase / Q000737:chemistry*; D009502:Neutrons; D011506:Proteins / Q000737:chemistry; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D012542:Scattering, Radiation; D012996:Solutions; D012997:Solvents / Q000737:chemistry; D013816:Thermodynamics; D014867:Water / Q000737:chemistry*
D011506:Proteins; D012996:Solutions; D012997:Solvents; D014867:Water; D012259:Ribonuclease, Pancreatic; D009113:Muramidase; D005990:Glycerol