title: Thermodynamic analysis of the nondenaturational conformational change of baker's yeast phosphoglycerate kinase at 24 degrees C.
abstract: A plot of the Gibbs free energy of unfolding vs. temperature is calculated for baker's yeast phosphoglycerate kinase in 150 mM sodium phosphate (pH=7.0) from a combination of reversible differential scanning calorimetry measurements and isothermal guanidine hydrochloride titrations. The stability curve reveals the existence of two stable, folded conformers with an abrupt conformational transition occurring at 24 degrees C. The transition state thermodynamics for the low- to high-temperature conformational change are calculated from slow-scan-rate differential scanning calorimetry measurements where it is found that the free energy barrier for the conversion is 90 kJ/mol and the transition state possesses a significant unfolding quality. This analysis also confirms a nondenaturational conformational transition at 24 degrees C. The data therefore suggest that X-ray structures obtained from crystals grown below this temperature may differ considerably from the physiological structure and that the two conformers are not readily interconverted.
authors.affiliation: Department of Chemistry and Physics, Texas Woman's University, Bell Avenue, Denton, TX 76204, USA.
authors.forename: Opral
authors.initials: O
authors.lastname: Ijeoma
authors.forename: Heather N
authors.initials: HN
authors.lastname: Hollowell
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Bodnar
authors.forename: B Mark
authors.initials: BM
authors.lastname: Britt
mesh_terms: D002152:Calorimetry, Differential Scanning; D004795:Enzyme Stability; D019791:Guanidine; D010735:Phosphoglycerate Kinase / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D013696:Temperature; D013816:Thermodynamics
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D010735:Phosphoglycerate Kinase; D019791:Guanidine