Direct determination of phosphate sugars in biological material by (1)H high-resolution magic-angle-spinning NMR spectroscopy.
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.
Departments of Radiology and Clinical Research, University of Bern, Murtenstrasse 35, 3008, Bern, Switzerland. gaelle.diserens@insel.ch.|Graduate School for Cellular and Biomedical Sciences, University of Bern, Freiestrasse 1, 3012, Bern, Switzerland. gaelle.diserens@insel.ch.
Gaëlle
G
Diserens
Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012, Bern, Switzerland.
Martina
M
Vermathen
Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012, Bern, Switzerland.
Ilche
I
Gjuroski
University Institute of Clinical Chemistry, Bern University Hospital, F/INO, 3010, Bern, Switzerland.
Sandra
S
Eggimann
Department of Clinical Veterinary Medicine, University of Bern, Länggassstrasse 124, 3001, Bern, Switzerland.
Christina
C
Precht
Departments of Radiology and Clinical Research, University of Bern, Murtenstrasse 35, 3008, Bern, Switzerland.
Chris
C
Boesch
Departments of Radiology and Clinical Research, University of Bern, Murtenstrasse 35, 3008, Bern, Switzerland.
Peter
P
Vermathen
D000818:Animals; D045744:Cell Line, Tumor; D006801:Humans; D018482:Muscle, Skeletal / Q000737:chemistry*; D009374:Neoplasms, Experimental / Q000737:chemistry*; D066244:Proton Magnetic Resonance Spectroscopy / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D013403:Sugar Phosphates / Q000032:analysis* / Q000737:chemistry*
D013403:Sugar Phosphates
1H HR-MAS NMR; Glucose-phosphates; Nucleotide sugars; UDP-X