title: Conformational stability of the Escherichia coli HPr protein: test of the linear extrapolation method and a thermodynamic characterization of cold denaturation.
abstract: The conformational stability of the histidine-containing phosphocarrier protein (HPr) from Escherichia coli has been determined using a combination of thermal unfolding and urea denaturation experiments. The analysis of the denaturation data provides a measure of the changes in conformational free energy, enthalpy, entropy, and heat capacity that accompany the equilibrium folding of HPr over a wide range of temperature and urea concentrations. In moderate concentrations of urea, HPr undergoes both high- and low-temperature unfolding, allowing for a reliable determination of the change in heat capacity for the conformational transition. The data are consistent with the linear free energy relationship commonly employed to analyze protein denaturation data, even over a relatively large temperature and urea concentration range. Furthermore, we find that a temperature-independent delta Cp is adequate to describe HPr stability over the accessible temperature range. Finally, our data allow us to evaluate the energetics of the urea-protein interaction. For HPr, the changes in excess enthalpy and entropy of the denaturant-protein interaction(s) make only minor contributions to the observed delta H and delta S terms, presumably due in some part to the small size of the HPr protein.
authors.affiliation: Department of Biochemistry and Biophysics, Texas A&M University, College Station 77843, USA.
authors.forename: E M
authors.initials: EM
authors.lastname: Nicholson
authors.forename: J M
authors.initials: JM
authors.lastname: Scholtz
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D002942:Circular Dichroism; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D013696:Temperature; D013816:Thermodynamics; D014508:Urea
chemical_list: D001426:Bacterial Proteins; D014508:Urea; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C030182:phosphocarrier protein HPr