Extraction and Measurement the Activities of Cytosolic Phosphoenolpyruvate Carboxykinase (PEPCK) and Plastidic NADP-dependent Malic Enzyme (ME) on Tomato (Solanum lycopersicum)
Sonia Osorio,
José Vallarino,
Marek Szecowka,
Shai Ufaz,
Vered Tzin,
Ruthie Angelovici,
Gad Galili,
Fayezeh Aarabi
Affiliations
Sonia Osorio
Department of Molecular Biology and Biochemistry, Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora” – University of Malaga- Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Malaga, Spain
José Vallarino
Department of Molecular Biology and Biochemistry, Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora” – University of Malaga- Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Malaga, Spain
Marek Szecowka
Central Metabolism Research Group, Max-Planck-Institut für Molekulare Planzenphysiologie, Potsdam-Golm, Germany
Shai Ufaz
Department of Plant Sciences, Weizmann Institute of Science, Rehovot, Israel
Vered Tzin
Department of Plant Sciences, Weizmann Institute of Science, Rehovot, Israel
Ruthie Angelovici
Department of Plant Sciences, Weizmann Institute of Science, Rehovot, Israel
Gad Galili
Department of Plant Sciences, Weizmann Institute of Science, Rehovot, Israel
Fayezeh Aarabi
Central Metabolism Research Group, Max-Planck-Institut für Molekulare Planzenphysiologie, Potsdam-Golm, Potsdam-Golm
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+.
