Binding of 3-phosphoglycerate leads to both activation and stabilisation of ADP-glucose pyrophosphorylase from apple leaves
Rui Zhou A and Lailiang Cheng A BA Department of Horticulture, Cornell University, Ithaca, NY 14853, USA.
B Corresponding author. Email: lc89@cornell.edu
Functional Plant Biology 32(9) 839-848 https://doi.org/10.1071/FP05055
Submitted: 21 March 2005 Accepted: 4 May 2005 Published: 26 August 2005
Abstract
Apple leaf ADP-glucose pyrophosphorylase was purified 1436-fold to apparent homogeneity with a specific activity of 58.9 units mg–1. The enzyme was activated by 3-phosphoglycerate (PGA) and inhibited by inorganic phosphate (Pi) in the ADPG synthesis direction. In the pyrophosphorolytic direction, however, high concentrations of PGA (> 2.5 mm) inhibited the enzyme activity. The enzyme was resistant to thermal inactivation with a T0.5 (temperature at which 50% of the enzyme activity is lost after 5 min incubation) of 52°C. Incubation with 2 mm PGA or 2 mm Pi increased T0.5 to 68°C. Incubation with 2 mm dithiothreitol (DTT) decreased T0.5 to 42°C, whereas inclusion of 2 mm PGA in the DTT incubation maintained T0.5 at 52°C. DTT-induced decrease in thermal stability was accompanied by monomerisation of the small subunits. Presence of PGA in the DTT incubation did not alter the monomerisation of the small subunits of the enzyme induced by DTT. These findings indicate that binding of PGA renders apple leaf AGPase with a conformation that is not only more efficient in catalysis but also more stable to heat treatment. The physiological significance of the protective effect of PGA on thermal inactivation of AGPase is discussed.
Keywords: ADP-glucose pyrophosphorylase, allosteric regulation, apple, inorganic phosphate, thermal stability, 3-phosphoglycerate.
Acknowledgments
We thank Dr Randy Wayne and Dr Robert Turgeon for their assistance in the purification and assay of the enzyme. Dr Thomas Okita kindly provided the antibody of potato tuber AGPase.
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