Electron Spray Ionization Mass Spectrometry And 2D P-31 Nmr For Monitoring O-18/O-16 Isotope Exchange And Turnover Rates Of Metabolic Oligophosphates

Date
2012Author
Nemutlu, Emirhan
Juranic, Nenad
Zhang, Song
Ward, Lawrence E.
Dutta, Tumpa
Nair, K. Sreekumaran
Terzic, Andre
Macura, Slobodan
Dzeja, Petras P.
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A new method was here developed for the determination of O-18-labeling ratios in metabolic oligophosphates, such as ATP, at different phosphoryl moieties (alpha-, beta-, and gamma-ATP) using sensitive and rapid electrospray ionization mass spectrometry (ESI-MS). The ESI-MS-based method for monitoring of O-18/O-16 exchange was validated with gas chromatography-mass spectrometry and 2D P-31 NMR correlation spectroscopy, the current standard methods in labeling studies. Significant correlation was found between isotopomer selective 2D P-31 NMR spectroscopy and isotopomer less selective ESI-MS method. Results demonstrate that ESI-MS provides a robust analytical platform for simultaneous determination of levels, O-18-labeling kinetics and turnover rates of alpha-, beta-, and gamma-phosphoryls in ATP molecule. Such method is advantageous for large scale dynamic phosphometabolomic profiling of metabolic networks and acquiring information on the status of probed cellular energetic system.