Abstract
Photorespiration is an essential metabolic process in plants occurring via the oxygenase reaction of RuBisCO. In order to understand this process, it is essential to determine the amounts of intermediates involved. For this purpose we combined mass spectrometry-based approaches and the use of authentic standards for the quantification of photorespiratory intermediates. Here we describe protocols for the extraction and quantification of 2-phosphoglycolate (2PG) by LC-MS/MS and serine, glycine, glycolate, hydroxypyruvate, glyoxylate, and glycerate by GC-MS.
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References
Arrivault S, Guenther M, Ivakov A et al (2009) Use of reverse-phase liquid chromatography, linked to tandem mass spectrometry, to profile the Calvin cycle and other metabolic intermediates in Arabidopsis rosettes at different carbon dioxide concentrations. Plant J 59(5):824–839
Arrivault S, Guenther M, Fry SC et al (2015) Synthesis and use of stable-isotope-labeled internal standards for quantification of phosphorylated metabolites by LC-MS/MS. Anal Chem 87(13):6896–6904
Lisec J, Schauer N, Kopka J et al (2006) Gas chromatography mass spectrometry-based metabolite profiling in plants. Nat Protoc 1(1):387–396
Boldt R, Edner C, Kolukisaoglu U et al (2005) D-GLYCERATE 3-KINASE, the last unknown enzyme in the photorespiratory cycle in Arabidopsis, belongs to a novel kinase family. Plant Cell 17(8):2413–2420
Pick TR, Brautigam A, Schulz MA et al (2013) PLGG1, a plastidic glycolate glycerate transporter, is required for photorespiration and defines a unique class of metabolite transporters. Proc Natl Acad Sci U S A 110(8):3185–3190
Heise R, Arrivault S, Szecowka M et al (2014) Flux profiling of photosynthetic carbon metabolism in intact plants. Nat Protoc 9(8):1803–1824
Luedemann A, von Malotky L, Erban A et al (2012) TagFinder: preprocessing software for the fingerprinting and the profiling of gas chromatography-mass spectrometry based metabolome analyses. Methods Mol Biol 860:255–286
Acknowledgments
Work from S.A. and T.O. was supported by the European Commission FP7 collaborative project 3 to 4 (Contract No. 289582).
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Arrivault, S., Obata, T. (2017). Quantification of Photorespiratory Intermediates by Mass Spectrometry-Based Approaches. In: Fernie, A., Bauwe, H., Weber, A. (eds) Photorespiration. Methods in Molecular Biology, vol 1653. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7225-8_7
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DOI: https://doi.org/10.1007/978-1-4939-7225-8_7
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7224-1
Online ISBN: 978-1-4939-7225-8
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