Electron spray ionization mass spectrometry and 2D P-31 NMR for monitoring O-18/O-16 isotope exchange and turnover rates of metabolic oligophosphates
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, cilt.403, sa.3, ss.697-706, 2012 (SCI-Expanded, Scopus)
- 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
- Açık Arşiv Koleksiyonu: AVESİS Açık Erişim Koleksiyonu
- 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.