adenosine-5--o-(3-thiotriphosphate) and glucose-1-phosphate

Substrate binding properties of the large (LS) and small (SS) subunits of potato tuber ADP-glucose pyrophosphorylase were investigated by using isothermal titration calorimetry. Our results clearly show that the wild type heterotetramer (S(WT)L(WT)) possesses two distinct types of ATP binding sites, whereas the homotetrameric LS and SS variant forms only exhibited properties of one of the two binding sites. The wild type enzyme also exhibited significantly increased affinity to this substrate compared to the homotetrameric enzyme forms. No stable binding was evident for the second substrate, glucose-1-phosphate, in the presence or absence of ATPγS suggesting that interaction of glucose-1-phosphate is dependent on hydrolysis of ATP and supports the Theorell-Chance bi bi reaction mechanism.

Topics: Adenosine Triphosphate; Binding Sites; Binding, Competitive; Blotting, Western; Calorimetry; Glucose-1-Phosphate Adenylyltransferase; Glucosephosphates; Kinetics; Models, Molecular; Molecular Structure; Plant Proteins; Plant Tubers; Protein Binding; Protein Multimerization; Protein Structure, Tertiary; Protein Subunits; Solanum tuberosum; Substrate Specificity; Thermodynamics