Insights into Neuronal Cell Metabolism Using NMR Spectroscopy: Uridyl Diphosphate <i>N</i>‐Acetyl‐Glucosamine as a Unique Metabolic Marker

Gallinger, Anika; Biet, Thorsten; Pellerin, Luc; Peters, Thomas

doi:10.1002/ange.201104836

Cited by 3 publications

(21 citation statements)

References 23 publications

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“…(Bruker Daltonics). Natural isotopic ions (X) were as follows: 13 Fig. 2A, panel 0 h) and thus disregarded in the analysis.…”

Section: Methodsmentioning

confidence: 99%

“…2A (panels 0.5 h, 2 h, and 6 h), was a result of the incorporation of 13 C 6 -hexose and its metabolites ( 13 C 5 -ribose and 13 C 2 -acetyl moieties) into UDP-GlcNAc, in combination with the natural occurrence of X 0 -X 2 mass isotopomers (supplemental Table S1). For example, the isotopic ion of m/z ϭ 614 contains 13 13 C 5 -ribose labeled motifs were calculated from the isotopic ions detected at m/z ϭ 611, 613, 617, and 619. The intensities of the 13 C 2 -acetyl residues were calculated from the isotopic ions detected at m/z ϭ 608, 613, 614, and 619.…”

Section: Methodsmentioning

confidence: 99%

“…The possible replacement patterns of the structural motifs in CMPNeuAc were also calculated as the combination of isotopic ions labeled at 13 C 6 -hexose, 13 C 5 -ribose, 13 C 2 -acetyl, and 13 C 3 -pyruvate (Supplemental Table 1). To estimate the replacement ratio of the 13 C 6 -hexose motif, we calculated the combined intensities of isotopic ions detected at m/z ϭ 619, 622, 624, 626, 627, and 629.…”

Section: Methodsmentioning

confidence: 99%

“…To estimate the replacement ratio of the 13 C 6 -hexose motif, we calculated the combined intensities of isotopic ions detected at m/z ϭ 619, 622, 624, 626, 627, and 629. The ion at m/z ϭ 621.1 had two possible patterns of labeling ( 13 C 6 -hexose ϩ 13 C 2 -acetyl and 13 C 5 -ribose ϩ 13 C 3 -pyruvate), and hence we represented the replacement ratios of 13 C 6 -hexose as possible ranges (maximum: including this peak; minimum: not including this peak).…”

Section: Methodsmentioning

confidence: 99%

“…For example, the use of nuclear magnetic resonance to obtain isotopomer signals of metabolically labeled molecules has been applied to trace the flux in glycolysis and fatty acid metabolism (13). An approach based on the mass isotopomers of labeled metabolites with 13 C 6 -glucose has been developed to monitor the UDP-GlcNAc synthetic pathway (13)(14)(15).…”

mentioning

confidence: 99%

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Mass Isotopomer Analysis of Metabolically Labeled Nucleotide Sugars and N- and O-Glycans for Tracing Nucleotide Sugar Metabolisms

Nakajima

Ito

Ohtsubo

et al. 2013

Molecular & Cellular Proteomics

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Nucleotide sugars are the donor substrates of various glycosyltransferases, and an important building block in N-and O-glycan biosynthesis. Their intercellular concentrations are regulated by cellular metabolic states including diseases such as cancer and diabetes. To investigate the fate of UDP-GlcNAc, we developed a tracing method for UDP-GlcNAc synthesis and use, and GlcNAc utilization using 13 C 6 -glucose and 13 C 2 -glucosamine, respectively, followed by the analysis of mass isotopomers using LC-MS.Metabolic labeling of cultured cells with 13 C 6 -glucose and the analysis of isotopomers of UDP-HexNAc (UDPGlcNAc plus UDP-GalNAc) and CMP-NeuAc revealed the relative contributions of metabolic pathways leading to UDP-GlcNAc synthesis and use. In pancreatic insulinoma cells, the labeling efficiency of a 13 C 6 -glucose motif in CMP-NeuAc was lower compared with that in hepatoma cells.Using 13 C 2 -glucosamine, the diversity of the labeling efficiency was observed in each sugar residue of N-and O-glycans on the basis of isotopomer analysis. In the insulinoma cells, the low labeling efficiencies were found for sialic acids as well as tri-and tetra-sialo N-glycans, whereas asialo N-glycans were found to be abundant. Essentially no significant difference in secreted hyaluronic acids was found among hepatoma and insulinoma cell lines. This indicates that metabolic flows are responsible for the low sialylation in the insulinoma cells. Our strategy should be useful for systematically tracing each stage of cellular GlcNAc metabolism. Molecular & Cellular Proteomics

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“…(Bruker Daltonics). Natural isotopic ions (X) were as follows: 13 Fig. 2A, panel 0 h) and thus disregarded in the analysis.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Mass Isotopomer Analysis of Metabolically Labeled Nucleotide Sugars and N- and O-Glycans for Tracing Nucleotide Sugar Metabolisms

Nakajima

Ito

Ohtsubo

et al. 2013

Molecular & Cellular Proteomics

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NMR Metabolomics for Stem Cell type discrimination

Castiglione

Ferro

Mavroudakis

et al. 2017

Sci Rep

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Cell metabolism is a key determinant factor for the pluripotency and fate commitment of Stem Cells (SCs) during development, ageing, pathological onset and progression. We derived and cultured selected subpopulations of rodent fetal, postnatal, adult Neural SCs (NSCs) and postnatal glial progenitors, Olfactory Ensheathing Cells (OECs), respectively from the subventricular zone (SVZ) and the olfactory bulb (OB). Cell lysates were analyzed by proton Nuclear Magnetic Resonance (1H-NMR) spectroscopy leading to metabolites identification and quantitation. Subsequent multivariate analysis of NMR data by Principal Component Analysis (PCA), and Partial Least Square Discriminant Analysis (PLS-DA) allowed data reduction and cluster analysis. This strategy ensures the definition of specific features in the metabolic content of phenotypically similar SCs sharing a common developmental origin. The metabolic fingerprints for selective metabolites or for the whole spectra demonstrated enhanced peculiarities among cell types. The key result of our work is a neat divergence between OECs and the remaining NSC cells. We also show that statistically significant differences for selective metabolites characterizes NSCs of different ages. Finally, the retrived metabolome in cell cultures correlates to the physiological SC features, thus allowing an integrated bioengineering approach for biologic fingerprints able to dissect the (neural) SC molecular specificities.

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The Effect of Antitumor Glycosides on Glioma Cells and Tissues as Studied by Proton HR-MAS NMR Spectroscopy

et al. 2013

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The effect of the treatment with glycolipid derivatives on the metabolic profile of intact glioma cells and tumor tissues, investigated using proton high resolution magic angle spinning (1H HR-MAS) nuclear magnetic resonance (NMR) spectroscopy, is reported here. Two compounds were used, a glycoside and its thioglycoside analogue, both showing anti-proliferative activity on glioma C6 cell cultures; however, only the thioglycoside exhibited antitumor activity in vivo. At the drug concentrations showing anti-proliferative activity in cell culture (20 and 40 µM), significant increases in choline containing metabolites were observed in the 1H NMR spectra of the same intact cells. In vivo experiments in nude mice bearing tumors derived from implanted C6 glioma cells, showed that reduction of tumor volume was associated with significant changes in the metabolic profile of the same intact tumor tissues; and were similar to those observed in cell culture. Specifically, the activity of the compounds is mainly associated with an increase in choline and phosphocholine, in both the cell cultures and tumoral tissues. Taurine, a metabolite that has been considered a biomarker of apoptosis, correlated with the reduction of tumor volume. Thus, the results indicate that the mode of action of the glycoside involves, at least in part, alteration of phospholipid metabolism, resulting in cell death.

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Insights into Neuronal Cell Metabolism Using NMR Spectroscopy: Uridyl Diphosphate N‐Acetyl‐Glucosamine as a Unique Metabolic Marker

Cited by 3 publications

References 23 publications

Mass Isotopomer Analysis of Metabolically Labeled Nucleotide Sugars and N- and O-Glycans for Tracing Nucleotide Sugar Metabolisms

Mass Isotopomer Analysis of Metabolically Labeled Nucleotide Sugars and N- and O-Glycans for Tracing Nucleotide Sugar Metabolisms

NMR Metabolomics for Stem Cell type discrimination

The Effect of Antitumor Glycosides on Glioma Cells and Tissues as Studied by Proton HR-MAS NMR Spectroscopy

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