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; Alisdair Fernie

doi:10.21769/BioProtoc.1122

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, Alisdair Fernie

The French Associates Institute for Agriculture and Biotechnology of Drylands

Research output: Contribution to journal › Article › peer-review

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+.

Original language	English
Pages (from-to)	http://test.bio-protocol.org/e1122
Journal	Bio-protocol
Volume	4
DOIs	https://doi.org/10.21769/BioProtoc.1122
State	Published - 1 Jan 2014

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10.21769/BioProtoc.1122

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Osorio, S., Vallarino, J., Szecowka, M., Ufaz, S., Tzin, V., Angelovici, R., Galili, G., & Fernie, A. (2014). Extraction and Measurement the Activities of Cytosolic Phosphoenolpyruvate Carboxykinase (PEPCK) and Plastidic NADP-dependent Malic Enzyme (ME) on Tomato (Solanum lycopersicum). Bio-protocol, 4, http://test.bio-protocol.org/e1122. https://doi.org/10.21769/BioProtoc.1122

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title = "Extraction and Measurement the Activities of Cytosolic Phosphoenolpyruvate Carboxykinase (PEPCK) and Plastidic NADP-dependent Malic Enzyme (ME) on Tomato (Solanum lycopersicum)",

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+.",

author = "Sonia Osorio and Jos{\'e} Vallarino and Marek Szecowka and Shai Ufaz and Vered Tzin and Ruthie Angelovici and Gad Galili and Alisdair Fernie",

year = "2014",

month = jan,

day = "1",

doi = "10.21769/BioProtoc.1122",

language = "English",

volume = "4",

pages = "http://test.bio--protocol.org/e1122",

journal = "Bio-protocol",

issn = "2331-8325",

publisher = "Bio-protocol LLC",

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Osorio, S, Vallarino, J, Szecowka, M, Ufaz, S, Tzin, V, Angelovici, R, Galili, G & Fernie, A 2014, 'Extraction and Measurement the Activities of Cytosolic Phosphoenolpyruvate Carboxykinase (PEPCK) and Plastidic NADP-dependent Malic Enzyme (ME) on Tomato (Solanum lycopersicum)', Bio-protocol, vol. 4, pp. http://test.bio-protocol.org/e1122. https://doi.org/10.21769/BioProtoc.1122

Extraction and Measurement the Activities of Cytosolic Phosphoenolpyruvate Carboxykinase (PEPCK) and Plastidic NADP-dependent Malic Enzyme (ME) on Tomato (Solanum lycopersicum). / Osorio, Sonia; Vallarino, José; Szecowka, Marek et al.
In: Bio-protocol, Vol. 4, 01.01.2014, p. http://test.bio-protocol.org/e1122.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Extraction and Measurement the Activities of Cytosolic Phosphoenolpyruvate Carboxykinase (PEPCK) and Plastidic NADP-dependent Malic Enzyme (ME) on Tomato (Solanum lycopersicum)

AU - Osorio, Sonia

AU - Vallarino, José

AU - Szecowka, Marek

AU - Ufaz, Shai

AU - Tzin, Vered

AU - Angelovici, Ruthie

AU - Galili, Gad

AU - Fernie, Alisdair

PY - 2014/1/1

Y1 - 2014/1/1

N2 - 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+.

AB - 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+.

U2 - 10.21769/BioProtoc.1122

DO - 10.21769/BioProtoc.1122

M3 - Article

SN - 2331-8325

VL - 4

SP - http://test.bio-protocol.org/e1122

JO - Bio-protocol

JF - Bio-protocol

ER -

Extraction and Measurement the Activities of Cytosolic Phosphoenolpyruvate Carboxykinase (PEPCK) and Plastidic NADP-dependent Malic Enzyme (ME) on Tomato (Solanum lycopersicum)

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