Abstract
Glutamate plays a central role in amino acid metabolism, in particular, in aminotransferase reactions leading to formation of many other proteinogenic and nonproteinogenic amino acids. In stress conditions, glutamate can be either metabolized to γ-aminobutyric acid (GABA) by glutamate decarboxylase which initiates a GABA shunt bypassing several reactions of the tricarboxylic acid cycle or converted to 2-oxoglutarate by glutamate dehydrogenase. Both reactions direct protein carbon to respiration but also link glutamate metabolism to other cellular pathways, resulting in the regulation of redox level and pH balance. Assays for determination of activities of glutamate dehydrogenase and of the GABA shunt enzymes as the markers of stress response is described in this chapter. These assays are important in the studies of the strategy of biochemical adaptation of plants to changing environmental conditions including elevated CO2, temperature increase, flooding, and other stresses.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Mano J (2012) Reactive carbonyl species: their production from lipid peroxides, action in environmental stress, and the detoxification mechanism. Plant Physiol Biochem 59:90–97
Kinnersley AM, Turano FJ (2000) γ-Aminobutyric acid (GABA) and plant responses to stress. Crit Rev Plant Sci 19:479–509
Popov VN, Eprintsev AT, Fedorin DN, Fomenko OY, Igamberdiev AU (2007) Role of transamination in the mobilization of respiratory substrates in germinating seeds of castor oil plants. Appl Biochem Microbiol 43:341–346
Fait A, Fromm H, Walter D, Galili G, Fernie AR (2008) Highway or byway: the metabolic role of the GABA shunt in plants. Trends Plant Sci 13:14–19
Khan MI, Nazir F, Asgher M, Per TS, Khan NA (2015) Selenium and sulfur influence ethylene formation and alleviate cadmium-induced oxidative stress by improving proline and glutathione production in wheat. J Plant Physiol 173:9–18
Yang H, Sun M, Lin S, Guo Y, Yang Y, Zhang T, Zhang J (2017) Transcriptome analysis of Crossostephium chinensis provides insight into the molecular basis of salinity stress responses. PLoS One 12:e0187124
Bown AW, Shelp BJ (1997) The metabolism and functions ofγ-aminobutyric acid. Plant Physiol 115:1–5
Michaeli S, Fait A, Lagor K, Nunes-Nesi A, Grillich N, Yellin A, Bar D, Khan M, Fernie AR, Turano FJ, Fromm H (2011) A mitochondrial GABA permease connects the GABA shunt and the TCA cycle, and is essential for normal carbon metabolism. Plant J 67:485–498
Renault H, El Amrani A, Berger A, Mouille G, Soubigou-Taconnat L, Bouchereau A, Deleu C (2013) γ-Aminobutyric acid transaminase deficiency impairs central carbon metabolism and leads to cell wall defects during salt stress in Arabidopsis roots. Plant Cell Environ 36:1009–1018
Steward FC, Thompson JF, Dent CE (1949) γ-Aminobutyric acid: a constituent of the potato tuber? Science 110:439–440
Corrales AR, Carrillo L, Lasierra P, Nebauer SG, Dominguez-Figueroa J, Renau-Morata B, Pollmann S, Granell A, Molina RV, Vicente-Carbajosa J, Medina J (2017) Multifaceted role of cycling DOF factor 3 (CDF3) in the regulation of flowering time and abiotic stress responses in Arabidopsis. Plant Cell Environ 40:748–764
Signorelli S, Dans PD, Coitiño EL, Borsani O, Monza J (2015) Connecting proline and γ-aminobutyric acid in stressed plants through non-enzymatic reactions. PLoS One 10:e0115349
Bowsher CG, Lacey AE, Hanke GT, Clarkson DT, Saker LR, Stulen I, Emes MJ (2007) The effect of Glc6P uptake and its subsequent oxidation within pea root plastids on nitrite reduction and glutamate synthesis. J Exp Bot 58:1109–1118
Breitkreuz KE, Allan WL, Van Cauwenberghe OR, Jakobs C, Talibi D, Andre B, Shelp BJ (2003) A novel gamma-hydroxybutyrate dehydrogenase: identification and expression of an Arabidopsis cDNA and potential role under oxygen deficiency. J Biol Chem 278:41552–41556
Busch KB, Fromm H (1999) Plant succinic semialdehyde dehydrogenase. Cloning, purification, localization in mitochondria, and regulation by adenine nucleotides. Plant Physiol 121:589–598
Busch KB, Piehler J, Fromm H (2000) Plant succinic semialdehyde dehydrogenase: dissection of nucleotide binding by surface plasmon resonance and fluorescence spectroscopy. Biochemistry 39:10110–10117
Andriamampandry C, Siffert J-C, Schmitt M, Garnier J-M, Staub A, Muller C, Gobaille S, Mark J, Maitre M (1998) Cloning of a rat brain succinic semialdehyde reductase involved in the synthesis of the neuromodulator gamma-hydroxybutyrate. Biochem J 334:43–50
Schaller M, Schaffhauser M, Sans N, Wermuth B (1999) Cloning and expression of succinic semialdehyde reductase from human brain. Identity with aflatoxin B1 aldehyde reductase. Eur J Biochem 265:1056–1060
Allan WL, Simpson JP, Clark SM, Shelp BJ (2008) Gamma-hydroxybutyrate accumulation in Arabidopsis and tobacco plants is a general response to abiotic stress: putative regulation by redox balance and glyoxylate reductase isoforms. J Exp Bot 59:2555–2564
Drew MC (1997) Oxygen deficiency and root metabolism: injury and acclimation under hypoxia and anoxia. Annu Rev Plant Physiol Plant Mol Biol 48:223–250
Allan WL, Clark SM, Hoover GJ, Shelp BJ (2009) Role of plant glyoxylate reductases during stress: a hypothesis. Biochem J 423:15–22
Tsai K-J, Lin C-Y, Ting C-Y, Shih M-C (2016) Ethylene-regulated glutamate dehydrogenase fine-tunes metabolism during anoxia-reoxygenation. Plant Physiol 172:1548–1562
Fontaine JX, Tercé-Laforgue T, Armengaud P, Clément G, Renou JP, Pelletier S, Catterou M, Azzopardi M, Gibon Y, Lea PJ, Hirel B, Dubois F (2012) Characterization of a NADH-dependent glutamate dehydrogenase mutant of Arabidopsis demonstrates the key role of this enzyme in root carbon and nitrogen metabolism. Plant Cell 24:4044–4065
Melo-Oliveira R, Oliveira IC, Coruzzi GM (1996) Arabidopsis mutant analysis and gene regulation define a nonredundant role for glutamate dehydrogenase in nitrogen assimilation. Proc Natl Acad Sci U S A 93:4718–4723
Loyola-Vargas VM, de Jimenez ES (1984) Differential role of glutamate dehydrogenase in nitrogen metabolism of maize tissues. Plant Physiol 76:536–540
Glevarec G, Bouton S, Jaspard E, Riou MT, Cliquet JB, Suzuki A, Limami AM (2004) Respective roles of the glutamine synthetase/glutamate synthase cycle and glutamate dehydrogenase in ammonium and amino acid metabolism during germination and post-germinative growth in the model legume Medicago truncatula. Planta 219:286–297
Bhatnagar P, Glasheen BM, Bains SK, Long SL, Minocha R, Walter C, Minocha SC (2001) Transgenic manipulation of the metabolism of polyamines in poplar cells. Plant Physiol 125:2139–2153
Hiraga K, Ueno Y, Oda K (2008) Glutamate decarboxylase from Lactobacillus brevis: activation by ammonium sulfate. Biosci Biotechnol Biochem 72:1299–1306
Huang Y, Su L, Wu J (2016) Pyridoxine supplementation improves the activity of recombinant glutamate decarboxylase and the enzymatic production of gamma-aminobutyric acid. PLoS One 11:e0157466
Park JY, Jeong SJ, Kim JH (2014) Characterization of a glutamate decarboxylase (GAD) gene from Lactobacillus zymae. Biotechnol Lett 36:1791–1799
Hampe CS, Hammerle LP, Falorni A, Robertson J, Lernmark A (2001) Site-directed mutagenesis of K396R of the 65 kDa glutamic acid decarboxylase active site obliterates enzyme activity but not antibody binding. FEBS Lett 488:185–189
Nonaka S, Someya T, Zhou S, Takayama M, Nakamura K, Ezura H (2017) An Agrobacterium tumefaciens strain with gamma-aminobutyric acid transaminase activity shows an enhanced genetic transformation ability in plants. Sci Rep 7:42649
Saito N, Robert M, Kochi H, Matsuo G, Kakazu Y, Soga T, Tomita M (2009) Metabolite profiling reveals YihU as a novel hydroxybutyrate dehydrogenase for alternative succinic semialdehyde metabolism in Escherichia coli. J Biol Chem 284:16442–16451
Narsai R, Howell KA, Carroll A, Ivanova A, Millar AH, Whelan J (2009) Defining core metabolic and transcriptomic responses to oxygen availability in rice embryos and young seedlings. Plant Physiol 151:306–322
Igamberdiev AU, Kleczkowski LA (2018) The glycerate and phosphorylated pathways of serine synthesis in plants: the branches of plant glycolysis linking carbon and nitrogen metabolism. Front Plant Sci 9:318
Kleczkowski LA, Edwards GE (1989) Identification of hydroxypyruvate and glyoxylate reductases in maize leaves. Plant Physiol 91:278–286
Acknowledgments
Funding: This work was supported by the grant 19-14-00150 of the Russian Science Foundation.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Eprintsev, A.T., Selivanova, N.V., Igamberdiev, A.U. (2020). Enzymatic Conversions of Glutamate and γ-Aminobutyric Acid as Indicators of Plant Stress Response. In: Gupta, K. (eds) Nitrogen Metabolism in Plants. Methods in Molecular Biology, vol 2057. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9790-9_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9790-9_7
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9789-3
Online ISBN: 978-1-4939-9790-9
eBook Packages: Springer Protocols