Limits...
2-Deoxy-2-fluoro-d-glucose metabolism in Arabidopsis thaliana.

Fatangare A, Paetz C, Saluz H, Svatoš A - Front Plant Sci (2015)

Bottom Line: We fed FDG to leaf tissue and analyzed leaf extracts using MS and NMR.Glycolysis and starch degradation seemed to be the important pathways for FDG metabolism.We showed that FDG metabolism in plants is considerably different than animal cells and goes beyond FDG-phosphate as previously presumed.

View Article: PubMed Central - PubMed

Affiliation: Mass Spectrometry/Proteomics Research Group, Max Planck Institute for Chemical Ecology Jena, Germany.

ABSTRACT
2-Deoxy-2-fluoro-d-glucose (FDG) is glucose analog routinely used in clinical and animal radiotracer studies to trace glucose uptake but it has rarely been used in plants. Previous studies analyzed FDG translocation and distribution pattern in plants and proposed that FDG could be used as a tracer for photoassimilates in plants. Elucidating FDG metabolism in plants is a crucial aspect for establishing its application as a radiotracer in plant imaging. Here, we describe the metabolic fate of FDG in the model plant species Arabidopsis thaliana. We fed FDG to leaf tissue and analyzed leaf extracts using MS and NMR. On the basis of exact mono-isotopic masses, MS/MS fragmentation, and NMR data, we identified 2-deoxy-2-fluoro-gluconic acid, FDG-6-phosphate, 2-deoxy-2-fluoro-maltose, and uridine-diphosphate-FDG as four major end products of FDG metabolism. Glycolysis and starch degradation seemed to be the important pathways for FDG metabolism. We showed that FDG metabolism in plants is considerably different than animal cells and goes beyond FDG-phosphate as previously presumed.

No MeSH data available.


Related in: MedlinePlus

Assignment of the fluorinated disaccharide. (A) Detail of the 1H-13C HSQC spectrum from the fraction containing the fluorinated disaccharide m/z 343.1051. Characteristic C-F and H-F couplings are given. Two different shifts (δc 99.3/99.5) for C-1′ appear depending from the configuration of the FDG part. The F2-Projection shows the selective TOCSY spectra for the α/β-FDG part. (B) Key correlations used for the structure elucidation of the fluorinated disaccharide m/z 343.1051. Blue arrows indicate 1H-13C HMBC correlations from the position 1 of the respective sugar units. The red parts of the structure indicate for neighboring positions probed by selective COSY experiments. The green double tipped arrow shows the NOE evidence for the α(14) junction between the two sugar units.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4630959&req=5

Figure 4: Assignment of the fluorinated disaccharide. (A) Detail of the 1H-13C HSQC spectrum from the fraction containing the fluorinated disaccharide m/z 343.1051. Characteristic C-F and H-F couplings are given. Two different shifts (δc 99.3/99.5) for C-1′ appear depending from the configuration of the FDG part. The F2-Projection shows the selective TOCSY spectra for the α/β-FDG part. (B) Key correlations used for the structure elucidation of the fluorinated disaccharide m/z 343.1051. Blue arrows indicate 1H-13C HMBC correlations from the position 1 of the respective sugar units. The red parts of the structure indicate for neighboring positions probed by selective COSY experiments. The green double tipped arrow shows the NOE evidence for the α(14) junction between the two sugar units.

Mentions: The 19F-NMR spectrum of the semi-purified sample showed two signals at δF − 198.26 (β-F-maltose) and δF − 198.50 (α-F-maltose), revealing this compound to be the second most abundant metabolite formed in A. thaliana after FDG administration. The 1H-1H dqfCOSY spectrum showed characteristic crosspeaks (Supplementary Figure 10). The 3J-coupling partners of H-1α/β show the large characteristic split caused by 2JHF coupling. Similar to α/β-FDG-6-P, two signals at δH 5.31 (d, 3JHH = 3.8 Hz, H-1α) and δH 4.78 (dd, 3JHH = 7.8/3JHF = 2.2 Hz, H-1β), respectively, represent the anomeric position 1 of the parent FDG structure. A signal overlapping with H-1α was assigned to H-1′. Again, selective TOCSY spectra have been employed to reduce signal overlap from impurities (Supplementary Figure 11). The resulting spectra were used as projections for 2D experiments (1H-1H dqfCOSY, 1H-13C HSQC, and 1H-13C HMBC). Characteristic signals and coupling constants are shown in a section of the 1H-13C HSQC spectrum (Figure 4A). Structure elucidation was based on information gathered from 1H-13C-HMBC and a series of selective 1H-1H COSY and selective 1H-1H NOESY spectra (Figure 4B). The structures with chemical shifts are summarized in Supplementary Figures 12A,B.


2-Deoxy-2-fluoro-d-glucose metabolism in Arabidopsis thaliana.

Fatangare A, Paetz C, Saluz H, Svatoš A - Front Plant Sci (2015)

Assignment of the fluorinated disaccharide. (A) Detail of the 1H-13C HSQC spectrum from the fraction containing the fluorinated disaccharide m/z 343.1051. Characteristic C-F and H-F couplings are given. Two different shifts (δc 99.3/99.5) for C-1′ appear depending from the configuration of the FDG part. The F2-Projection shows the selective TOCSY spectra for the α/β-FDG part. (B) Key correlations used for the structure elucidation of the fluorinated disaccharide m/z 343.1051. Blue arrows indicate 1H-13C HMBC correlations from the position 1 of the respective sugar units. The red parts of the structure indicate for neighboring positions probed by selective COSY experiments. The green double tipped arrow shows the NOE evidence for the α(14) junction between the two sugar units.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4630959&req=5

Figure 4: Assignment of the fluorinated disaccharide. (A) Detail of the 1H-13C HSQC spectrum from the fraction containing the fluorinated disaccharide m/z 343.1051. Characteristic C-F and H-F couplings are given. Two different shifts (δc 99.3/99.5) for C-1′ appear depending from the configuration of the FDG part. The F2-Projection shows the selective TOCSY spectra for the α/β-FDG part. (B) Key correlations used for the structure elucidation of the fluorinated disaccharide m/z 343.1051. Blue arrows indicate 1H-13C HMBC correlations from the position 1 of the respective sugar units. The red parts of the structure indicate for neighboring positions probed by selective COSY experiments. The green double tipped arrow shows the NOE evidence for the α(14) junction between the two sugar units.
Mentions: The 19F-NMR spectrum of the semi-purified sample showed two signals at δF − 198.26 (β-F-maltose) and δF − 198.50 (α-F-maltose), revealing this compound to be the second most abundant metabolite formed in A. thaliana after FDG administration. The 1H-1H dqfCOSY spectrum showed characteristic crosspeaks (Supplementary Figure 10). The 3J-coupling partners of H-1α/β show the large characteristic split caused by 2JHF coupling. Similar to α/β-FDG-6-P, two signals at δH 5.31 (d, 3JHH = 3.8 Hz, H-1α) and δH 4.78 (dd, 3JHH = 7.8/3JHF = 2.2 Hz, H-1β), respectively, represent the anomeric position 1 of the parent FDG structure. A signal overlapping with H-1α was assigned to H-1′. Again, selective TOCSY spectra have been employed to reduce signal overlap from impurities (Supplementary Figure 11). The resulting spectra were used as projections for 2D experiments (1H-1H dqfCOSY, 1H-13C HSQC, and 1H-13C HMBC). Characteristic signals and coupling constants are shown in a section of the 1H-13C HSQC spectrum (Figure 4A). Structure elucidation was based on information gathered from 1H-13C-HMBC and a series of selective 1H-1H COSY and selective 1H-1H NOESY spectra (Figure 4B). The structures with chemical shifts are summarized in Supplementary Figures 12A,B.

Bottom Line: We fed FDG to leaf tissue and analyzed leaf extracts using MS and NMR.Glycolysis and starch degradation seemed to be the important pathways for FDG metabolism.We showed that FDG metabolism in plants is considerably different than animal cells and goes beyond FDG-phosphate as previously presumed.

View Article: PubMed Central - PubMed

Affiliation: Mass Spectrometry/Proteomics Research Group, Max Planck Institute for Chemical Ecology Jena, Germany.

ABSTRACT
2-Deoxy-2-fluoro-d-glucose (FDG) is glucose analog routinely used in clinical and animal radiotracer studies to trace glucose uptake but it has rarely been used in plants. Previous studies analyzed FDG translocation and distribution pattern in plants and proposed that FDG could be used as a tracer for photoassimilates in plants. Elucidating FDG metabolism in plants is a crucial aspect for establishing its application as a radiotracer in plant imaging. Here, we describe the metabolic fate of FDG in the model plant species Arabidopsis thaliana. We fed FDG to leaf tissue and analyzed leaf extracts using MS and NMR. On the basis of exact mono-isotopic masses, MS/MS fragmentation, and NMR data, we identified 2-deoxy-2-fluoro-gluconic acid, FDG-6-phosphate, 2-deoxy-2-fluoro-maltose, and uridine-diphosphate-FDG as four major end products of FDG metabolism. Glycolysis and starch degradation seemed to be the important pathways for FDG metabolism. We showed that FDG metabolism in plants is considerably different than animal cells and goes beyond FDG-phosphate as previously presumed.

No MeSH data available.


Related in: MedlinePlus