Glycerol Decreases the Volume and Compressibility of Protein Interior

• Published in: Biochemistry (Easton) 1996-02, Vol.35 (7), p.2061-2066
• Main Authors: Priev, A, Almagor, A, Yedgar, S, Gavish, B
• Format: Article
• Language: English
• Subjects: Amino Acids - chemistry; Carbohydrates - chemistry; Glycerol - chemistry; Hydrogen Bonding; Proteins - chemistry; Thermodynamics
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi951842r

The addition of hydrogen-bonded cosolvents to aqueous solutions of proteins is known to modify both thermodynamic and dynamic properties of the proteins in a variety of ways. Previous studies suggest that glycerol reduces the free volume and compressibility of proteins. However, there is no directly measured evidence for that. We have measured the apparent specific volume (V) and adiabatic compressibility (K) of a number of proteins, sugars, and amino acids in water and in 30% glycerol at pH 7.4 and 30 °C. The values of V and K in water and their changes induced by glycerol were extrapolated to the limit of infinite solute size. The main results were the following:  (a) glycerol decreases V and K of proteins, but increases it for amino acids; (b) the V and K values of the protein interior in water were found to be 0.784 ± 0.026 mL/g and (12.8 ± 2.5) × 10-6 mL/g·atm, where the glycerol reduces these values by 8 and 32%, respectively; (c) the coefficient of adiabatic compressibility of the structural component of proteins affected by the glycerol is estimated to be (50 ± 10) × 10-6 atm-1, which is comparable to that of water. We propose that the glycerol induces a release of the so-called “lubricant” water, which maintains conformational flexibility by keeping apart neighboring segments of the polypeptide chain. This is expected to lead to the collapsing of the voids containing this water as well as to increase intramolecular bonding, which explains the observed effect.