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Isothermal acid-titration calorimetry for evaluating the pH dependence of protein stability.
Biophys Chem. 2004 May 01; 109(2):229-49.BC

Abstract

A new method, which can be called as isothermal acid-titration calorimetry (IATC), was proposed for evaluating the enthalpy of protein molecules as a function of pH using isothermal titration calorimetry (ITC). This measurement was used to analyze the acid-denaturation of bovine ribonuclease A. The enthalpy change by acid-denaturation of this protein was estimated as 310 kJ/mol at pH 2.8 and 40 degrees C. This value agreed well with the enthalpy change obtained by differential scanning calorimetry. The midpoint pH and proton binding-number difference observed by IATC agreed well with those of the acid transition of the three-dimensional structure monitored by circular dichroism spectrometry. The van't Hoff enthalpy of the transition was derived from the temperature dependence of the midpoint pH and the proton binding-number difference. It agreed well with the calorimetric enthalpy change directly observed by IATC, strongly indicating that there was no stable intermediate state during the acid transition of this protein.

Authors+Show Affiliations

Nakamura S
Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka 940-2188, Japan.
Kidokoro S
No affiliation info available

MeSH

AlgorithmsAnimalsCalorimetryCattleCircular DichroismHot TemperatureHydrogen-Ion ConcentrationLeast-Squares AnalysisProtein DenaturationRibonuclease, PancreaticTemperatureThermodynamicsTitrimetry

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15110942

Citation

Nakamura, Shigeyoshi, and Shun-ichi Kidokoro. "Isothermal Acid-titration Calorimetry for Evaluating the pH Dependence of Protein Stability." Biophysical Chemistry, vol. 109, no. 2, 2004, pp. 229-49.
Nakamura S, Kidokoro S. Isothermal acid-titration calorimetry for evaluating the pH dependence of protein stability. Biophys Chem. 2004;109(2):229-49.
Nakamura, S., & Kidokoro, S. (2004). Isothermal acid-titration calorimetry for evaluating the pH dependence of protein stability. Biophysical Chemistry, 109(2), 229-49.
Nakamura S, Kidokoro S. Isothermal Acid-titration Calorimetry for Evaluating the pH Dependence of Protein Stability. Biophys Chem. 2004 May 1;109(2):229-49. PubMed PMID: 15110942.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isothermal acid-titration calorimetry for evaluating the pH dependence of protein stability. AU - Nakamura,Shigeyoshi, AU - Kidokoro,Shun-ichi, PY - 2003/10/02/received PY - 2003/11/19/revised PY - 2003/11/20/accepted PY - 2004/4/28/pubmed PY - 2005/10/22/medline PY - 2004/4/28/entrez SP - 229 EP - 49 JF - Biophysical chemistry JO - Biophys Chem VL - 109 IS - 2 N2 - A new method, which can be called as isothermal acid-titration calorimetry (IATC), was proposed for evaluating the enthalpy of protein molecules as a function of pH using isothermal titration calorimetry (ITC). This measurement was used to analyze the acid-denaturation of bovine ribonuclease A. The enthalpy change by acid-denaturation of this protein was estimated as 310 kJ/mol at pH 2.8 and 40 degrees C. This value agreed well with the enthalpy change obtained by differential scanning calorimetry. The midpoint pH and proton binding-number difference observed by IATC agreed well with those of the acid transition of the three-dimensional structure monitored by circular dichroism spectrometry. The van't Hoff enthalpy of the transition was derived from the temperature dependence of the midpoint pH and the proton binding-number difference. It agreed well with the calorimetric enthalpy change directly observed by IATC, strongly indicating that there was no stable intermediate state during the acid transition of this protein. SN - 0301-4622 UR - https://www.unboundmedicine.com/medline/citation/15110942/Isothermal_acid_titration_calorimetry_for_evaluating_the_pH_dependence_of_protein_stability_ DB - PRIME DP - Unbound Medicine ER -