Measurement of Tricarboxylic Acid Cycle Enzyme Activities in Plants
Mitochondria are vital cytoplasmic organelle of eukaryotic cells responsible for oxidative energy metabolism and the synthesis of intermediates utilized in various other metabolic pathways. The functions of mitochondrion are the oxidation of organic acids by the tricarboxylic acid (TCA) cycle and the synthesis of ATP by the oxidative phosphorylation in the mitochondrial electron transport chain. The TCA cycle is composed by a set of enzymes that are essential for optimal functioning of the primary carbon metabolism in plants. The activity of each TCA cycle enzyme in plants may vary according to cell type, plant tissue, stage of plant development, and the environment. Here, we describe current methods used for the determination of the TCA cycle enzyme activities in different plant tissues.
Key wordsActivation assays Mitochondrial enzyme activity Tricarboxylic acid (TCA) cycle enzyme assays
Financial support was provided by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq- to W.L.A.), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), and Max Planck Society to A.N.N. and W.L.A. Research fellowships granted by CNPq to A.N.N. and W.L.A. as well as scholarship granted by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) to R.P.O.G. are also gratefully acknowledged.
Conflict of interest: The authors declare that they have no conflict of interest.
- 2.Araújo WL, Nunes-Nesi A, Nikoloski Z et al (2012) Metabolic control and regulation of the tricarboxylic acid cycle in photosynthetic and heterotrophic plant tissues. Plant Cell Environ 35:1–21. doi: 10.1111/j.1365-3040.2011.02332.x
- 6.Daloso DM, Müller K, Obata T et al (2015) Thioredoxin, a master regulator of the tricarboxylic acid cycle in plant mitochondria. Proc Natl Acad Sci 112. doi: 10.1073/pnas.1424840112
- 8.Araújo WL, Nunes-Nesi A, Trenkamp S et al (2008) Inhibition of 2-oxoglutarate dehydrogenase in potato tuber suggests the enzyme is limiting for respiration and confirms its importance in nitrogen assimilation. Plant Physiol 148:1782–1796. doi: 10.1104/pp.108.126219 CrossRefPubMedPubMedCentralGoogle Scholar
- 13.Randall DD, Miernyk J (2012) 10 the mitochondrial pyruvate dehydrogenase complex. Enzymes of primary. Metabolism 3:175Google Scholar
- 14.Gibon Y, Blaesing OE, Hannemann J et al (2004) A robot-based platform to measure multiple enzyme activities in arabidopsis using a set of cycling assays: comparison of changes of enzyme activities and transcript levels during diurnal cycles and in prolonged darkness. Plant Cell 16:3304–3325. doi: 10.1105/tpc.104.025973 CrossRefPubMedPubMedCentralGoogle Scholar
- 16.Araújo WL, Nunes-Nesi A, Osorio S et al (2011) Antisense inhibition of the iron-sulphur subunit of succinate dehydrogenase enhances photosynthesis and growth in tomato via an organic acid–mediated effect on stomatal aperture. Plant Cell 23:600–627. doi: 10.1105/tpc.110.081224 CrossRefPubMedPubMedCentralGoogle Scholar