Isotopic profiling of C-13-labeled biological samples by two-dimensional heteronuclear J-resolved nuclear magnetic resonance spectroscopy

Edern Cahoreau (1, 2, 3) , Lindsay Peyriga (1, 2) , Jane Hubert (1, 2, 4) , Frederic Bringaud (4) , Stéphane Massou (1, 2) , Jean-Charles Portais (2, 1)

1 UT3 - Université Toulouse III - Paul Sabatier
2 LISBP - Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés
3 TBI-MetaToul - MetaToul FluxoMet
4 CRMSB - Centre de résonance magnétique des systèmes biologiques

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 C-13-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 C-13 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. (C) 2012 Elsevier Inc. All rights reserved.

Mots clés: H-1 NMR; Isotopic profiling; Carbon labeling experiment; C-13 enrichment; 2D NMR; J-resolved spectroscopy; METABOLIC FLUX ANALYSIS; QUANTITATIVE-ANALYSIS; LABELING EXPERIMENTS; MASS-SPECTROMETRY; ESCHERICHIA-COLI; NMR-SPECTROSCOPY; C-13; STABILITY; GLUTAMATE; MIXTURES

Domaines: Sciences du Vivant [q-bio]

Edern Cahoreau, Lindsay Peyriga, Jane Hubert, Frederic Bringaud, Stéphane Massou, et al.. Isotopic profiling of C-13-labeled biological samples by two-dimensional heteronuclear J-resolved nuclear magnetic resonance spectroscopy. Analytical Biochemistry, 2012, 427 (2), pp.158 - 163. ⟨10.1016/j.ab.2012.05.021⟩. ⟨hal-01268291⟩