Direct determination of phosphate sugars in biological material by (1)H high-resolution magic-angle-spinning NMR spectroscopy

Anal Bioanal Chem. 2016 Aug;408(20):5651-6. doi: 10.1007/s00216-016-9671-0. Epub 2016 Jun 6.

Abstract

The study aim was to unambiguously assign nucleotide sugars, mainly UDP-X that are known to be important in glycosylation processes as sugar donors, and glucose-phosphates that are important intermediate metabolites for storage and transfer of energy directly in spectra of intact cells, as well as in skeletal muscle biopsies by (1)H high-resolution magic-angle-spinning (HR-MAS) NMR. The results demonstrate that sugar phosphates can be determined quickly and non-destructively in cells and biopsies by HR-MAS, which may prove valuable considering the importance of phosphate sugars in cell metabolism for nucleic acid synthesis. As proof of principle, an example of phosphate-sugar reaction and degradation kinetics after unfreezing the sample is shown for a cardiac muscle, suggesting the possibility to follow by HR-MAS NMR some metabolic pathways. Graphical abstract Glucose-phosphate sugars (Glc-1P and Glc-6P) detected in muscle by 1H HR-MAS NMR.

Keywords: 1H HR-MAS NMR; Glucose-phosphates; Nucleotide sugars; UDP-X.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Humans
  • Muscle, Skeletal / chemistry*
  • Neoplasms, Experimental / chemistry*
  • Proton Magnetic Resonance Spectroscopy / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sheep
  • Sugar Phosphates / analysis*
  • Sugar Phosphates / chemistry*

Substances

  • Sugar Phosphates