Analysis of individual purine and pyrimidine nucleoside di- and triphosphates and other cellular metabolites in PCA extracts by using multinuclear high resolution NMR spectroscopy.
This work demonstrates that individual purine and pyrimidine NDP and NTP can be assigned in high resolution 31P NMR spectra from tissue extracts. To the best of our knowledge, it is shown for the first time that ATP, GTP, UTP, CTP, and the corresponding diphosphates can be quantitated in cell extracts without using HPLC or other biochemical methods. This work provides the basis for further optimization of nucleotide quantitation by 31P NMR spectroscopy, and for a full assessment of this method. Furthermore, a new technique was developed for 1H, 31P, and 13C NMR signal assignment and quantitation in cell extracts by using the same external reference capillary for all three nuclei. This allows for efficient, quantitative, multinuclear NMR spectroscopy without extract contamination by standard material.
Centre de Résonance Magnétique Biologique et Médicale, URA CNRS 1186, Faculté de Médecine, Marseille, France.
N W
NW
Lutz
N
N
Yahi
J
J
Fantini
P J
PJ
Cozzone
D000244:Adenosine Diphosphate / Q000032:analysis; D000255:Adenosine Triphosphate / Q000032:analysis; D003565:Cytidine Diphosphate / Q000032:analysis; D003570:Cytidine Triphosphate / Q000032:analysis; D006153:Guanosine Diphosphate / Q000032:analysis; D006160:Guanosine Triphosphate / Q000032:analysis; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011685:Purine Nucleotides / Q000032:analysis*; D011742:Pyrimidine Nucleotides / Q000032:analysis*; D014530:Uridine Diphosphate / Q000032:analysis; D014544:Uridine Triphosphate / Q000032:analysis
D011685:Purine Nucleotides; D011742:Pyrimidine Nucleotides; D006153:Guanosine Diphosphate; D014530:Uridine Diphosphate; D000244:Adenosine Diphosphate; D003565:Cytidine Diphosphate; D003570:Cytidine Triphosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D014544:Uridine Triphosphate