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• W2023204648 abstract "The Ca(2+)-ATPase from sarcoplasmic reticulum (SR) membranes couples the Ca(2+) transport to ATP hydrolysis through phosphorylation in its cytoplasmic catalytic domain. Interactions between protein domains and the role of monomer-monomer interactions remain unclear. Here, we report a differential scanning calorimetric study of the thermal unfolding of this protein. In the pH range 6-8, thermal unfolding of the Ca(2+)-ATPase in glycogen phosphorylase-free SR membranes shows a major endothermic peak with a critical temperature midpoint ranging between 51 and 55 degrees C, depending on pH, Ca(2+), Mg(2+)-ADP and KCl concentrations. The enthalpy change of the overall unfolding process ranged between 250 and 300 kcal/mol of Ca(2+)-ATPase monomer. Thermal denaturation of the Ca(2+)-ATPase in SR membranes is well fitted to an irreversible process that can be rationalized in terms of a non-two state process, N (native)right harpoon over left harpoon I (intermediate)-->D (denatured). Thermodynamic analysis show that this protein has a compact structure, implying a tight structural interconnection between catalytic and Ca(2+) transport domains. The apparent cooperative unit, defined by the van 't Hoff enthalpy to the overall unfolding enthalpy ratio, increased from 1.1 at pH 6 to 1.8 at pH 8, showing that monomer-monomer interactions are stronger at weakly basic pH than at weakly acidic pH. While micromolar Ca(2+) concentrations had only a weak effect on the cooperativity of the unfolding process, this is clearly increased by millimolar Mg(2+)-ADP. In addition, high ionic strength lowered the apparent cooperative unit to approximately 1.0 in the pH range 6-8. Taken together, these results suggest that protein-protein interactions are altered by variables that modulate the catalytic activity of this enzyme." @default.
• W2023204648 created "2016-06-24" @default.
• W2023204648 creator A5012237274 @default.
• W2023204648 creator A5046252340 @default.
• W2023204648 date "1999-08-01" @default.
• W2023204648 modified "2023-09-28" @default.
• W2023204648 title "pH and ligand binding modulate the strength of protein–protein interactions in the Ca2+-ATPase from sarcoplasmic reticulum membranes" @default.
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• W2023204648 doi "https://doi.org/10.1016/s0005-2736(99)00101-7" @default.
• W2023204648 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/10446303" @default.
• W2023204648 hasPublicationYear "1999" @default.
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