Analysis of individual purine and pyrimidine nucleoside di- and triphosphates and other cellular metabolites in PCA extracts by using multinuclear high resolution NMR spectroscopy
✍ Scribed by Norbert W. Lutz; Nouara Yahi; Jacques Fantini; Patrick J. Cozzone
- Publisher
- John Wiley and Sons
- Year
- 1996
- Tongue
- English
- Weight
- 811 KB
- Volume
- 36
- Category
- Article
- ISSN
- 0740-3194
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✦ Synopsis
Abstract
This work demonstrates that individual purine and pyrimidine NDP and NTP can be assigned in high resolution ^31^P 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 ^31^P NMR spectroscopy, and for a full assessment of this method. Furthermore, a new technique was developed for ^1^H, ^31^P, and ^13^C 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.