Experimental measurement of the effective dielectric in the hydrophobic core of a protein

• Published in: Biophysical chemistry Vol. 64; no. 1; pp. 211 - 224
• Main Authors: García-Moreno, Bertrand E., Dwyer, John J., Gittis, Apostolos G., Lattman, Eaton E., Spencer, Daniel S., Stites, Wesley E.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 28-02-1997
• Subjects: Buried ionizable group; Dielectric inside a protein; Effective dielectric; Electrostatics; Hydration; p Ka in hydrophobic environment; Proteins - chemistry; Static Electricity; Buried ionizable group; Dielectric inside a protein; Effective dielectric; Hydration; p K a in hydrophobic environment; Electrostatics
• ISSN: 0301-4622; 1873-4200
• DOI: 10.1016/S0301-4622(96)02238-7

The dielectric inside a protein is a key physical determinant of the magnitude of electrostatic interactions in proteins. We have measured this dielectric phenomenologically, in terms of the dielectric that needs to be used with the Born equation in order to reproduce the observed p
K
a shifts induced by burial of an ionizable group in the hydrophobic core of a protein. Mutants of staphylococcal nuclease with a buried lysine residue at position 66 were engineered for this purpose. The p
K
a values of buried lysines were measured by difference potentiometry. The extent of coupling between the p
K
a and the global stability of the protein was evaluated by measuring p
K
a values in hyperstable forms of nuclease engineered to be 3.3 or 6.5 kcal mol
−1 more stable than the wild type. The crystallographic structure of one mutant was determined to describe the environment of the buried lysine. The dielectrics that were measured range from 10 to 12. Published p
K
a values of buried ionizable residues in other proteins were analyzed in a similar fashion and the dielectrics obtained from these values are consistent with the ones measured in nuclease. These results argue strongly against the prevalent use of dielectrics of 4 or lower to describe the dielectric effect inside a protein in structure-based calculations of electrostatic energies with continuum dielectric models.