A new method for the determination of specific 13C enrichment in phosphorylated [1-13C]glucose metabolites. 13C-coupled, 1H-decoupled 31P-NMR spectroscopy of tissue perchloric acid extracts.
A 31P-NMR method for the determination of 13C enrichment in phosphorylated [1-(13)C]glucose metabolites was developed by taking advantage of the 13C satellites detectable for 31P-NMR signals of metabolites such as UDP-hexoses, UDP-N-acetylhexosamines and other phosphorylated compounds generated during glycolysis and subsequent anabolism. HT-29 cells were incubated in culture medium containing 4.5 g/l [1-(13)C]glucose for 24 h prior to cell extraction, and high-resolution 31P-NMR spectra were acquired from perchloric acid extracts. Since glucose and its phosphorylated products are key metabolites for many different metabolic processes, this method may be very helpful for studying specific metabolic pathways involving phosphorylated glucose metabolites.
Lutz
N W
NW
Centre de Résonance Magnétique Biologique et Médicale, URA CNRS 1186, Faculté de Médecine, Marseille, France.
Yahi
N
N
Fantini
J
J
Cozzone
P J
PJ
D002247:Carbon Isotopes; D002457:Cell Extracts; D003110:Colonic Neoplasms; D003470:Culture Media; D005947:Glucose / Q000378:metabolism*; D006019:Glycolysis; D006595:Hexosamines / Q000378:metabolism; D006601:Hexoses / Q000378:metabolism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D010472:Perchlorates; D010710:Phosphates / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D010766:Phosphorylation; D011687:Purines / Q000378:metabolism; D011743:Pyrimidines / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000032:analysis; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000032:analysis
D002247:Carbon Isotopes; D002457:Cell Extracts; D003470:Culture Media; D006595:Hexosamines; D006601:Hexoses; D010472:Perchlorates; D010710:Phosphates; D010743:Phospholipids; D011687:Purines; D011743:Pyrimidines; D014537:Uridine Diphosphate N-Acetylglucosamine; D014536:Uridine Diphosphate N-Acetylgalactosamine; D005947:Glucose