Isotopic profiling of ¹³C-labeled biological samples by two-dimensional heteronuclear J-resolved nuclear magnetic resonance spectroscopy.
The use of two-dimensional heteronuclear J-resolved (2D H-JRES) nuclear magnetic resonance (NMR) spectroscopy for fast and reliable measurement of isotopic patterns from ¹³C-enriched compounds resulting from carbon labeling experiments was evaluated. Its use with biological samples of increasing complexity showed that 2D H-JRES spectroscopy is suitable for high-throughput isotopic profiling of any kind of labeled samples. Moreover, the method enabled accurate quantification of ¹³C enrichments and, thus, can be used for metabolic flux analysis. The excellent trade-off between reduced experimental time and the number of measurable isotopic data makes 2D H-JRES NMR a promising approach for high-throughput flux analysis of samples of intermediate complexity.
Université de Toulouse, INSA, UPS, INP, LISBP, F-31077 Toulouse, France.
Edern
E
Cahoreau
Lindsay
L
Peyriga
Jane
J
Hubert
Frédéric
F
Bringaud
Stéphane
S
Massou
Jean-Charles
JC
Portais
D000596:Amino Acids / Q000032:analysis*; D002247:Carbon Isotopes; D045424:Complex Mixtures / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry*; D057166:High-Throughput Screening Assays; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D014346:Trypanosoma brucei brucei / Q000737:chemistry*
D000596:Amino Acids; D002247:Carbon Isotopes; D045424:Complex Mixtures