Direct determination of thiol pKa by isothermal titration microcalorimetry.
Knowledge of the acid dissociation constant (pKa) of a molecule is a critical step toward understanding its structure and reactivity. Current methods for pKa measurement, including electrochemical, spectroscopic, and spectrophotometric titrations, have proven to be useful but also have significant limitations. To overcome these limitations, we report the use of isothermal titration calorimetry (ITC) as a new method for pKa determination. We demonstrate by the measurement of the pKa values for free cysteine, glutathione, and a cysteine residue in a protein that this method is rapid and accurate.
Department of Biochemistry and Biophysics, University of Rochester, Rochester, New York 14642, USA.
Stephen G
SG
Tajc
Blanton S
BS
Tolbert
Ravi
R
Basavappa
Benjamin L
BL
Miller
D001665:Binding Sites; D002151:Calorimetry / Q000379:methods*; D003545:Cysteine / Q000737:chemistry; D005978:Glutathione / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D017186:Titrimetry; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry
D013438:Sulfhydryl Compounds; C105156:UBE2C protein, human; D044763:Ubiquitin-Conjugating Enzymes; D005978:Glutathione; D003545:Cysteine