Abstract
Mass spectrometry (MS) in combination with isotope labelling experiments is widely used for investigations of metabolism and other biological processes. Quantitative applications—e.g., 13C metabolic flux analysis—require correction of raw MS data (isotopic clusters) for the contribution of all naturally abundant isotopes. This chapter describes how to perform such correction using the software IsoCor. This flexible, user-friendly software can be used to exploit any isotopic tracer, from well-known (13C, 15N, 18O, etc.) to unusual (57Fe, 77Se, etc.) isotopes. It also provides options—e.g., correction for the isotopic purity of the tracer—to improve the accuracy of quantitative isotopic studies, and allows automated correction of large datasets that can be collected with modern MS methods.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Nicolas C, Kiefer P et al (2007) Response of the central metabolism of Escherichia coli to modified expression of the gene encoding the glucose-6-phosphate dehydrogenase. FEBS Lett 581(20): 3771–3776
Perrenoud A, Sauer U (2005) Impact of global transcriptional regulation by ArcA, ArcB, Cra, Crp, Cya, Fnr, and Mlc on glucose catabolism in Escherichia coli. J Bacteriol 187(9):3171–3179
Kiefer P, Nicolas C et al (2007) Determination of carbon labeling distribution of intracellular metabolites from single fragment ions by ion chromatography tandem mass spectrometry. Anal Biochem 360(2):182–188
Rosman KJR, Taylor PDP (1998) Isotopic compositions of the elements 1997. Pure Appl Chem 70(1):217–235
van Winden WA, Wittmann C et al (2002) Correcting mass isotopomer distributions for naturally occurring isotopes. Biotechnol Bioeng 80(4):477–479
Millard P, Letisse F et al (2012) IsoCor: correcting MS data in isotope labeling experiments. Bioinformatics 28(9):1294–1296
Wittmann C (2007) Fluxome analysis using GC-MS. Microb Cell Fact 6:6
Wittmann C, Heinzle E (1999) Mass spectrometry for metabolic flux analysis. Biotechnol Bioeng 62(6):739–750
Rodriguez-Castrillon JA, Moldovan M et al (2008) Isotope pattern deconvolution as a tool to study iron metabolism in plants. Anal Bioanal Chem 390(2):579–590
Acknowledgements
The authors are thankful to members of MetaSys team (LISBP, Toulouse) for fruitful discussions. The PhD fellowship of P.M. was funded by the Institut National de la Recherche Agronomique (INRA) [Program CJS]. The continuous support of Région Midi-Pyrénées, the European Regional Development Fund (ERDF), the French Ministry for Higher Education and Research, the SICOVAL, and INRA is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Millard, P., Letisse, F., Sokol, S., Portais, JC. (2014). Correction of MS Data for Naturally Occurring Isotopes in Isotope Labelling Experiments. In: Krömer, J., Nielsen, L., Blank, L. (eds) Metabolic Flux Analysis. Methods in Molecular Biology, vol 1191. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1170-7_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1170-7_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1169-1
Online ISBN: 978-1-4939-1170-7
eBook Packages: Springer Protocols