Abstract
A recent study demonstrated that cytosolic phosphoenolpyruvate carboxykinase (PEPCK) and NADP-malic enzyme (NADP-ME) have an important role in malate metabolism during fruit ripening (Osorio et al., 2013). PEPCK catalyze the ATP-dependent decarboxylation of oxaloacetate (OAA) to phosphoenolpyruvate (PEP) and NADP-ME, the reversible conversion of malate and pyruvate. Here, we present the detailed protocols to measure PEPCK activity in carboxylation direction by following oxidation of NADH and to measure NADP-ME activity based upon the reduction of NADP+.
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
We acknowledge the excellent care of the plants by Helga Kulka (Max- Planck-Institut für Molekulare Planzenphysiologie) and Hanna Levanony (Weizmann Institute of Science). This protocol was adapted from Osorio et al. (2013).
References
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