31P-NMR studies of glucose and glutamine metabolism in cultured mammalian cells

Sri-Pathmanathan, Rohendra M.; Braddock, Peta; Brindle, Kevin M.

doi:10.1016/0167-4889(90)90184-f

Cited by 19 publications

(13 citation statements)

References 22 publications

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“…It should be noted that, upon switching from glucose to glutamine, there was a decrease in PCr to a new steady state that was -50% of the glucose-perifused baseline levels. Similar effects on PCr level were observed for glucose-versus glutamine-perifused HeLa cells (27). Because of this decrease, the data from the glutamine-perifused MMQ cells are more noisy.…”

Section: Characterization Of Mmq Cells Perifused With Glutarninesupporting

confidence: 75%

“…This slow decrease of NTP levels has been previously observed in both perifused human lymphocytes (25) and a hybridoma cell line (27). However, in other cell lines, stable NTP levels can be maintained for long periods of time (14,15,27). It is not clear why some cell lines can maintain stable NTP levels and others cannot.…”

Section: Characterization Of Perifused Mmq Cellsmentioning

confidence: 74%

“…1). Spectra of the MMQ cells acquired in 1 2 min are similar to the spectra of other perifused cell published previously (14, 15,25,27). The MMQcells contain a small amount of PCr when they are cultured in RPMI 1640 ( would be low.…”

Section: Characterization Of Perifused Mmq Cellsmentioning

confidence: 95%

“…Because the cells display a small resonance due to PCr under ordinary culture conditions, the addition of creatine presumably does not alter the metabolism of the cells greatly. Increasing the level of PCr in cultured cells has been accomplished successfully in a similar manner in perifused HeLa cells (27). In that study, the investigators were attempting to determine how glucose and glutamine would each affect the steady-state energy status of the perifused cells (27).…”

Section: Characterization Of Perifused Mmq Cellsmentioning

confidence: 98%

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³¹P NMR investigation of energy metabolism in perifused MMQ cells

Goger

Fernandez

et al. 1994

Magnetic Resonance in Med

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The MMQ cell line is a unique prolactin-secreting rat pituitary cell line. MMQ cells entrapped in agarose gel threads are metabolically active, as determined by the uptake and phosphorylation of creatine and the maintenance of high energy phosphates for over 15 h. Forskolin activates the catalytic subunit of adenylyl cyclase and, in MMQ cells, elevates the level of cAMP and stimulates prolactin secretion. 31P NMR spectroscopy was used to investigate the energy metabolism of the MMQ cells during stimulation by forskolin. The ability to measure small changes in the energy status of these cells was enhanced by increasing the PCr levels in the cells. Administration of forskolin to the perifused MMQ cells resulted in acute, reversible, and dose-dependent changes in the 31P NMR spectra of the cells within 12 to 24 min of the beginning of forskolin exposure. Several lines of evidence indicate that the changes observed in the MMQ cells are the composite result of the interaction of forskolin with adenylyl cyclase and the plasma membrane glucose transporter. Also, preincubation of the MMQ cells with the dopamine agonist, bromocriptine, attenuates the forskolin-stimulated decrease in the PCr resonance by approximately 50%. This attenuation indicates that the forskolin-stimulated changes in energy metabolism are probably related to the prolactin secretion process.

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Section: Characterization Of Mmq Cells Perifused With Glutarninesupporting

confidence: 75%

Section: Characterization Of Perifused Mmq Cellsmentioning

confidence: 74%

Section: Characterization Of Perifused Mmq Cellsmentioning

confidence: 95%

Section: Characterization Of Perifused Mmq Cellsmentioning

confidence: 98%

See 2 more Smart Citations

³¹P NMR investigation of energy metabolism in perifused MMQ cells

Goger

Fernandez

et al. 1994

Magnetic Resonance in Med

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“…Phosphate-activated glutaminase (PAG) enables the deamidation of glutamine to glutamate and in some cells has shown to be the rate-limiting step for glutamine utilization (Sri-Pathmanathan et al, 1990;Fitzpatrick et al, 1993). The activity of this enzyme decreased for BHK cells grown in glutamate-based media.…”

Section: Discussionmentioning

confidence: 99%

The adaptation of bhk cells to a non-ammoniagenic glutamate-based culture medium

Christie

Butler

1999

Biotechnol. Bioeng.

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Elevated glutamate dehydrogenase flux in glucose-deprived hybridoma and myeloma cells: Evidence from1H/15N NMR

Martinelle

Doverskog

Jacobsson

et al. 1998

Biotechnol. Bioeng.

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The glutamine metabolism was studied in glucose‐starved and glucose‐sufficient hybridoma and Sp2/0‐Ag14 myeloma cells. Glucose starvation was attained by cultivating the hybridoma cells with fructose instead of glucose, and the myeloma cells with a low initial glucose concentration which was rapidly exhausted. Glutamine used in the experiments was labeled with 15N, either in the amine or in the amide position. The fate of the label was monitored by 1H/15N NMR analysis of released 15NH 4+ and 15N‐alanine. Thus, NH 4+ formed via glutaminase (GLNase) could be distinguished from NH 4+ formed via glutamate dehydrogenase (GDH). In the glucose‐sufficient cells a small but measurable amount of 15NH 4+ released by GDH could be detected in both cell lines (0.75 and 0.31 μmole/106 cells for hybridoma and myeloma cells, respectively). The uptake of glutamine and the total production of NH 4+ was significantly increased in both fructose‐grown hybridoma and glucose‐starved myeloma cells, as compared to the glucose‐sufficient cells. The increased NH 4+ production was due to an increased throughput via GLNase (1.6 –1.9‐fold in the hybridoma, and 2.7‐fold in the myeloma cell line) and an even further increased metabolism via GDH (4.8–7.9‐fold in the hybridoma cells, and 3.1‐fold in the myeloma cells). The data indicate that both GLNase and GDH are down‐regulated when glucose is in excess, but up‐regulated in glucose‐starved cells. It was calculated that the maximum potential ATP production from glutamine could increase by 35–40 % in the fructose‐grown hybridoma cells, mainly due to the increased metabolism via GDH. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 60: 508–517, 1998.

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31P-NMR studies of glucose and glutamine metabolism in cultured mammalian cells

Cited by 19 publications

References 22 publications

³¹P NMR investigation of energy metabolism in perifused MMQ cells

³¹P NMR investigation of energy metabolism in perifused MMQ cells

The adaptation of bhk cells to a non-ammoniagenic glutamate-based culture medium

Elevated glutamate dehydrogenase flux in glucose-deprived hybridoma and myeloma cells: Evidence from1H/15N NMR

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31P-NMR studies of glucose and glutamine metabolism in cultured mammalian cells

Cited by 19 publications

References 22 publications

31P NMR investigation of energy metabolism in perifused MMQ cells

31P NMR investigation of energy metabolism in perifused MMQ cells

The adaptation of bhk cells to a non-ammoniagenic glutamate-based culture medium

Elevated glutamate dehydrogenase flux in glucose-deprived hybridoma and myeloma cells: Evidence from1H/15N NMR

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³¹P NMR investigation of energy metabolism in perifused MMQ cells

³¹P NMR investigation of energy metabolism in perifused MMQ cells