On the stabilizing action of protein denaturants: acetonitrile effect on stability of lysozyme in aqueous solutions.
Stability of hen lysozyme in the presence of acetonitrile (MeCN) at different pH values of the medium was studied by scanning microcalorimetry with a special emphasis on determination of reliable values of the denaturational heat capacity change. It was found that the temperature of denaturation decreases on addition of MeCN. However, the free energy extrapolation showed that below room temperature the thermodynamic stability increases at low concentrations of MeCN in spite of the general destabilizing effect at higher concentrations and temperatures. Charge-induced contribution to this stabilization was shown to be negligible (no pH-dependence was found); therefore, the most probable cause for the phenomenon is an increase of hydrophobic interactions at low temperatures in aqueous solutions containing small amounts of the organic additive. The difference in preferential solvation of native and denatured states of lysozyme was calculated from the stabilization free energy data. It was found that the change in preferential solvation strongly depends on the temperature in the water-rich region. At the higher MeCN content this dependence decreases until, at 0.06 mole fractions of MeCN, the difference in the preferential solvation between native and denatured lysozyme becomes independent of the temperature over a range of 60 K. The importance of taking into account non-ideality of a mixed solution, when analyzing preferential solvation phenomena was emphasized.
Kovrigin
E L
EL
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow, Russia.
Potekhin
S A
SA
D000097:Acetonitriles / Q000737:chemistry*; D000818:Animals; D002152:Calorimetry, Differential Scanning; D002645:Chickens; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D009113:Muramidase / Q000737:chemistry*; D011489:Protein Denaturation; D012996:Solutions / Q000737:chemistry; D013696:Temperature; D013816:Thermodynamics; D014867:Water / Q000737:chemistry
D000097:Acetonitriles; D012996:Solutions; D014867:Water; D009113:Muramidase; C032159:acetonitrile