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

Ben-Gurion University of the Negev

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",

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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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