Stabilizing effect of polyols is sensitive to inherent stability of protein.

In studies on polyol-mediated protein stabilization, the polyols are the preferred variable and less importance is given to the intrinsic properties of the protein used. We investigated the stabilizing effects of glycerol on three in vitro evolved lipase mutants with varying stabilities and also in a broad pH range of 3.3-12.1. Significant linear negative correlation between increment in stability due to glycerol and prior stability suggests that stabilizing effects of glycerol depend on the prior stability of the protein. Polar/nonpolar surface area and charge do not have a bearing on the stabilizing effects of glycerol.

[1]  N. Taulier,et al.  Compressibility of protein transitions. , 2002, Biochimica et biophysica acta.

[2]  R. Bhat,et al.  Thermal Stability of Proteins in Aqueous Polyol Solutions: Role of the Surface Tension of Water in the Stabilizing Effect of Polyols , 1998 .

[3]  P. Acharya,et al.  Crystallization and preliminary X-ray crystallographic investigations on several thermostable forms of a Bacillus subtilis lipase. , 2004, Acta crystallographica. Section D, Biological crystallography.

[4]  D. Erie,et al.  Interpreting the effects of small uncharged solutes on protein-folding equilibria. , 2001, Annual review of biophysics and biomolecular structure.

[5]  F. Ahmad,et al.  Stability of proteins in the presence of polyols estimated from their guanidinium chloride-induced transition curves at different pH values and 25 degrees C. , 2006, Biophysical chemistry.

[6]  Lenwood S. Heath,et al.  H++: a server for estimating pKas and adding missing hydrogens to macromolecules , 2005, Nucleic Acids Res..

[7]  D. W. Bolen,et al.  The peptide backbone plays a dominant role in protein stabilization by naturally occurring osmolytes. , 1995, Biochemistry.

[8]  N. Tolbert,et al.  Protein determination in membrane and lipoprotein samples: manual and automated procedures. , 1981, Methods in enzymology.

[9]  A. C. Ferreon,et al.  Metrics that differentiate the origins of osmolyte effects on protein stability: a test of the surface tension proposal. , 2006, Journal of molecular biology.

[10]  S. N. Timasheff,et al.  Protein hydration, thermodynamic binding, and preferential hydration. , 2002, Biochemistry.

[11]  F. Ahmad,et al.  Effect of polyol osmolytes on DeltaG(D), the Gibbs energy of stabilisation of proteins at different pH values. , 2005, Biophysical chemistry.

[12]  Rajan Sankaranarayanan,et al.  Thermostable Bacillus subtilis lipases: in vitro evolution and structural insight. , 2008, Journal of molecular biology.

[13]  N. Tolbert,et al.  A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. , 1978, Analytical biochemistry.

[14]  P. Acharya,et al.  Structural basis for the remarkable stability of Bacillus subtilis lipase (Lip A) at low pH. , 2008, Biochimica et biophysica acta.

[15]  Rajan Sankaranarayanan,et al.  Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase. , 2004, Journal of molecular biology.

[16]  B. Gavish,et al.  Reduction of protein volume and compressibility by macromolecular cosolvents: dependence on the cosolvent molecular weight. , 1998, Biochimica et biophysica acta.

[17]  K. Gekko,et al.  Mechanism of protein stabilization by glycerol: preferential hydration in glycerol-water mixtures. , 1981, Biochemistry.

[18]  Oleg V. Tsodikov,et al.  Novel computer program for fast exact calculation of accessible and molecular surface areas and average surface curvature , 2002, J. Comput. Chem..

[19]  C. Tanford.  Protein denaturation. , 1968, Advances in protein chemistry.

[20]  C. Tanford,et al.  Extension of the theory of linked functions to incorporate the effects of protein hydration. , 1969, Journal of molecular biology.

[21]  Emilia Bramanti,et al.  Effects of sucrose on the internal dynamics of azurin. , 2005, Biophysical journal.

[22]  D. Winzor,et al.  Measurement of thermodynamic nonideality arising from volume-exclusion interactions between proteins and polymers. , 1995, Biophysical chemistry.

[23]  A. Minton,et al.  Influence of excluded volume upon macromolecular structure and associations in 'crowded' media. , 1997, Current opinion in biotechnology.

[24]  R. Bhat,et al.  Why Is Trehalose an Exceptional Protein Stabilizer? , 2003, Journal of Biological Chemistry.

[25]  O. Berg,et al.  The influence of macromolecular crowding on thermodynamic activity: Solubility and dimerization constants for spherical and dumbbell‐shaped molecules in a hard‐sphere mixture , 1990, Biopolymers.

[26]  S. N. Timasheff.  Solvent stabilization of protein structure. , 1995, Methods in molecular biology.

[27]  J. Hermans,et al.  Polymer-protein interactions. Comparison of experiment and excluded volume theory. , 1983, The Journal of biological chemistry.

[28]  D. Allen,et al.  Osmolyte-induced changes in protein conformational equilibria. , 2000, Biopolymers.