Valence and anion binding of bovine ribonuclease A between pH 6 and 8
Abstract
Several studies have shown that divalent anion binding to ribonuclease A (RNase A) contributes to RNase A folding and stability. However, there are conflicting reports about whether chloride binds to or stabilizes RNase A. Two broad-zone experimental approaches, membrane-confined electrophoresis and analytical ultracentrifugation, were used to examine the electrostatic and electrohydrodynamic characteristics of aqueous solutions of bovine RNase A in the presence of 100 mM KCl and 10 mM Bis–Tris propane over a pH range of 6.00–8.00. The results of data analysis using a Debye–Hückel–Henry model, compared with expectations based on pKA values, are consistent with the binding of two chlorides by RNase A. The decreased protein valence resulting from anion binding contributes 2–3 kJ/mol to protein stabilization. This work demonstrates the utility of first-principle valence determinations to detect protein solution properties that might otherwise remain undetected.
Department
Molecular, Cellular and Biomedical Sciences
Publication Date
1-15-2005
Journal Title
Analytical Biochemistry
Publisher
Elsevier
Digital Object Identifier (DOI)
Document Type
Article
Recommended Citation
Moody TP, Kingsbury JS, Durant JA, Wilson TJ, Chase SF, Laue TM. (2005) "Valence and anion binding of bovine ribonuclease A between pH 6 and 8." Anal. Biochem. 336, 243-252.