Electron spray ionization mass spectrometry and 2D P-31 NMR for monitoring O-18/O-16 isotope exchange and turnover rates of metabolic oligophosphates

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NEMUTLU E., Juranic N., Zhang S., Ward L. E., Dutta T., Nair K. S., ...Daha Fazla

ANALYTICAL AND BIOANALYTICAL CHEMISTRY, cilt.403, sa.3, ss.697-706, 2012 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 403 Sayı: 3
  • Basım Tarihi: 2012
  • Doi Numarası: 10.1007/s00216-012-5899-5
  • Dergi Adı: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.697-706
  • Anahtar Kelimeler: O-18 isotope labeling, ESI-MS, P-31 NMR, ATP, Energy metabolism, Phosphotransfer networks, KINASE-CATALYZED PHOSPHOTRANSFER, ADENYLATE KINASE, PHOSPHORYL TRANSFER, CREATINE-KINASE, ENERGETIC COMMUNICATION, ADENOSINE-TRIPHOSPHATE, KNOCKOUT HEART, O-18, FLUX, NETWORKS
  • Hacettepe Üniversitesi Adresli: Evet

Özet

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.