Correlations Between the Charge of Proteins and the Number of Ionizable Groups They Incorporate: Studies Using Protein Charge Ladders, Capillary Electrophoresis, and Debye−Hückel Theory

Abstract

The values of electrophoretic mobility, μelectro, of bovine carbonic anhydrase II, human carbonic anhydrase II, cytochrome c, lysozyme, superoxide dismutase, ovalbumin, and derivatives of these proteins produced by partial neutralization of Lys e-NH3+ and/or Asp and Glu carboxyl groups were measured using capillary electrophoresis (CE). For derivatives of these proteins with the lowest overall values of net charge (either positive or negative), the values of μelectro and the values of charge measured by CE, ZCE, demonstrate a linear correlation with the number of charged groups, n, converted to neutral derivatives. For derivatives of these proteins with larger values of net charge, the values of μelectro and ZCE demonstrate a nonlinear correlation with n. Several observations made in this work suggest that shifts in the values of pka of the ionizable groups on these proteins likely contribute to the observed nonlinear correlation. Debye−Huckel theory was used to calculate values of electrostatic potential...