Thermodynamic and kinetic examination of protein stabilization by glycerol.
The effect of concentrated glycerol on the thermal transitions of chymotrypsinogen and ribonuclease has been examined by differential spectrophotometry at 293 and 287 mm, respectively. It was found that for both proteins addition of glycerol raises the transition temperature, the increase in Tm being greater for ribonuclease than for chymotrypsinogen. This increase in the free energy of denaturation appears to reflect primarily a decrease in the entropy change. Analysis in terms of the Wyman linkage equation shows that, for both proteins, the exclusion of glycerol from the protein domain increases on denaturation i.e., the chemical potential of glycerol becomes even more positive when the protein unfolds relative to the native structure. This provides the thermodynamic stabilization free energy. Results of the kinetic examination of the slow unfolding reaction are consistent with the concept that the preferential exclusion of glycerol is related, at least in part, to enhanced solvent ordering.
D000818:Animals; D002417:Cattle; D002919:Chymotrypsinogen; D004355:Drug Stability; D004720:Endonucleases / Q000378:metabolism*; D005990:Glycerol; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010179:Pancreas / Q000201:enzymology; D011489:Protein Denaturation; D012259:Ribonuclease, Pancreatic; D012260:Ribonucleases / Q000378:metabolism*; D013552:Swine; D013696:Temperature; D013816:Thermodynamics
D002919:Chymotrypsinogen; D004720:Endonucleases; D012260:Ribonucleases; D012259:Ribonuclease, Pancreatic; D005990:Glycerol