title: Scanning calorimetric study of the thermal unfolding of catabolite activator protein from Escherichia coli in the absence and presence of cyclic mononucleotides.
abstract: The thermal unfolding of the catabolite activator protein (CAP) of Escherichia coli and the complexes it forms with adenosine cyclic 3',5'-phosphate (cAMP) and guanosine cyclic 3',5'-phosphate (cGMP) was studied by high-sensitivity differential scanning calorimetry (DSC). The thermal denaturation of CAP at pH 7.00 gave an irreversible, symmetrical denaturation curve with a single peak. Distinctly different, more complex DSC curves were obtained for the thermal denaturation of the cAMP-protein and cGMP-protein complexes. The DSC data indicate intermolecular cooperation among CAP dimers, with the extent of oligomerization remaining unchanged during unfolding of the protein. The DSC curves for the thermal denaturation of the cAMP-protein complex and cGMP-protein complex have been resolved into three and two components, respectively, according to the model of independent two-state processes. Analysis of the DSC data suggests two and three independent domains for cGMP-protein and cAMP-protein complexes, respectively, with dissociation of mononucleotide occurring in the second component in both cases during protein denaturation. Furthermore, our studies indicate that the presence of either ligand alters the degree of oligomerization of CAP dimers, cAMP having a greater effect than cGMP.
authors.affiliation: Department of Chemistry, Yale University, New Haven, Connecticut 06511.
authors.forename: L R
authors.initials: LR
authors.lastname: Ghosaini
authors.forename: A M
authors.initials: AM
authors.lastname: Brown
authors.forename: J M
authors.initials: JM
authors.lastname: Sturtevant
mesh_terms: D002152:Calorimetry, Differential Scanning; D000242:Cyclic AMP / Q000378:metabolism*; D002373:Cyclic AMP Receptor Protein; D006152:Cyclic GMP / Q000378:metabolism*; D004926:Escherichia coli / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D011487:Protein Conformation
chemical_list: D002373:Cyclic AMP Receptor Protein; D000242:Cyclic AMP; D006152:Cyclic GMP