Effect of salt concentration on the pKa of acidic residues in lysozyme

Yoshito Abe, Tadashi Ueda, Hiroki Iwashita, Yoshio Hashimoto, Hiroyuki Motoshima, Yoshitugu Tanaka, Taiji Imoto

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Abstract

We determined the pKa values of acidic residues in hen lysozyme by comparing the pH dependency of stability between wild type and mutant lysozymes in which a negative charge is eliminated. In the comparison of the stability between wild type and a mutant lysozyme, the difference in pH titration curve between them could be expressed as a two-state process involving protonation of a single acidic residue. The results strongly indicated that the Aune and Tanford theory of protein denaturation [Aune, K.C. and Tanford, C. (1969) Biochemistry 8, 4579-4585] is applicable to protein stability in solution. On the other hand, the pKa values of acidic residues in the presence of low (5 mM) or high (400 mM) salt concentration were determined by means of two-dimensional NMR. We found that the pKa values obtained from the pH dependency of stability were close to those from the NMR experiment under the high salt condition. Moreover, by comparing pKa values at high salt and low salt concentrations, we could evaluate the dependency of two electrostatic interactions (salt bridge and charge-helix dipole interaction) on salt concentration.

Original languageEnglish
Pages (from-to)946-952
Number of pages7
JournalJournal of biochemistry
Volume118
Issue number5
DOIs
Publication statusPublished - Oct 1995

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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    Abe, Y., Ueda, T., Iwashita, H., Hashimoto, Y., Motoshima, H., Tanaka, Y., & Imoto, T. (1995). Effect of salt concentration on the pKa of acidic residues in lysozyme. Journal of biochemistry, 118(5), 946-952. https://doi.org/10.1093/jb/118.5.946