The effect of salt on protein chemical potential determined by ternary diffusion in aqueous solutions.
We use accurate thermodynamic derivatives extracted from high-precision measurements of the four volume-fixed diffusion coefficients in ternary solutions of lysozyme chloride in aqueous NaCl, NH4Cl, and KCl at pH 4.5 and 25 degrees C to (a) assess the relative contributions of the common-ion and nonideality effects to the protein chemical potential as a function of salt concentration, (b) compare the behavior of the protein chemical potential for the three salts, which we found to be consistent with the Hofmeister series, and (c) discuss our thermodynamic data in relation to the dependence of the protein solubility on salt concentration. The four diffusion coefficients are reported at 0.6 mM lysozyme chloride and 0.25, 0.5, 0.9, 1.2, and 1.5 M KCl and extend into the protein-supersaturated region. The chemical potential cross-derivatives are extracted from diffusion data using the Onsager reciprocal relation and the equality of molal cross-derivatives of solute chemical potentials. They are compared to those calculated previously from diffusion data for lysozyme in aqueous NaCl and NH4Cl. We estimate the effective charge on the diffusing lysozyme cation at the experimental concentrations. Our diffusion measurements on the three salts allowed us to analyze and interpret the four diffusion coefficients for charged proteins in the presence of 1:1 electrolytes. Our results may provide guidance to the understanding of protein crystallization.
Department of Chemistry, Texas Christian University, Fort Worth, Texas 76129, USA. O.Annunziata@tcu.edu
Onofrio
O
Annunziata
Luigi
L
Paduano
Arne J
AJ
Pearlstein
Donald G
DG
Miller
John G
JG
Albright
D000643:Ammonium Chloride / Q000737:chemistry*; D002712:Chlorides / Q000737:chemistry*; D004058:Diffusion; D009113:Muramidase / Q000737:chemistry*; D011189:Potassium Chloride / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D013816:Thermodynamics; D014867:Water / Q000737:chemistry
D002712:Chlorides; D012996:Solutions; D000643:Ammonium Chloride; D014867:Water; D012965:Sodium Chloride; D011189:Potassium Chloride; D009113:Muramidase