13C and 31P NMR investigation of effect of 6-Aminonicotinamide on metabolism of RIF-1 tumor cells in vitro

James C. Street, Umar Mahmood, Douglas Ballon, Alan A. Alfieri, Jason A. Koutcher

Research output: Contribution to journalArticle

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Abstract

The effect of 6-aminonicotinamide on the metabolism of RIF-1 tumor cells was investigated using 13C and 31P NMR spectroscopy. 6-Aminonicotinamide can be metabolized to 6-amino-NAD(P), a competitive inhibitor of NAD(P)-requiring processes. 40 μM 6-aminonicotinamide led to an inhibition of 6-phosphogluconate dehy-drogenase and an accumulation of 6-phosphogluconate. A subsequent accumulation of the 6-phosphogluconate precursor 6-phosphoglucono-δ-lactone was observed in the 13C NMR spectrum. These metabolites were shown to be intracellular, although a small amount of leakage of 6-phosphoglucono-δ-lactone occurred. The intracellular concentrations of 6-phosphogluconate and 6-phosphoglucono-δ-lactone were 1.9 ± 0.8 μmol/108 cells (±1 standard deviation) and 0.8 ± 0.4 μmol/108 cells, respectively, after 15 h. Glucose utilization and lactate production were significantly inhibited by 6-aminonicotinamide (both p < 0.05), indicating inhibition of glycolysis. 31P NMR data showed that phosphocreatine was significantly depleted in cells exposed to 6-aminonicotinamide (p < 0.05). Exposure of RIF-1 cells to 6-aminonicotinamide prior to 3- or 6-Gy x-irradiation induced a supra-additive cell kill, indicating that 6-aminonicotinamide is acting as a radiosensitizer. There was no effect of 6-aminonicotinamide alone or when the drug was given postradiation, suggesting that its mechanism of action may be by inhibition of radiation-induced repair.

Original languageEnglish (US)
Pages (from-to)4113-4119
Number of pages7
JournalJournal of Biological Chemistry
Volume271
Issue number8
StatePublished - Feb 23 1996
Externally publishedYes

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6-Aminonicotinamide
Metabolism
Tumors
Cells
Nuclear magnetic resonance
Neoplasms
Lactones
Phosphocreatine
In Vitro Techniques
Carbon-13 Magnetic Resonance Spectroscopy
Glycolysis
Metabolites
NAD
Nuclear magnetic resonance spectroscopy
Lactic Acid
Repair
Magnetic Resonance Spectroscopy
Irradiation
Radiation
Glucose

ASJC Scopus subject areas

  • Biochemistry

Cite this

Street, J. C., Mahmood, U., Ballon, D., Alfieri, A. A., & Koutcher, J. A. (1996). 13C and 31P NMR investigation of effect of 6-Aminonicotinamide on metabolism of RIF-1 tumor cells in vitro. Journal of Biological Chemistry, 271(8), 4113-4119.

13C and 31P NMR investigation of effect of 6-Aminonicotinamide on metabolism of RIF-1 tumor cells in vitro. / Street, James C.; Mahmood, Umar; Ballon, Douglas; Alfieri, Alan A.; Koutcher, Jason A.

In: Journal of Biological Chemistry, Vol. 271, No. 8, 23.02.1996, p. 4113-4119.

Research output: Contribution to journalArticle

Street, JC, Mahmood, U, Ballon, D, Alfieri, AA & Koutcher, JA 1996, '13C and 31P NMR investigation of effect of 6-Aminonicotinamide on metabolism of RIF-1 tumor cells in vitro', Journal of Biological Chemistry, vol. 271, no. 8, pp. 4113-4119.
Street, James C. ; Mahmood, Umar ; Ballon, Douglas ; Alfieri, Alan A. ; Koutcher, Jason A. / 13C and 31P NMR investigation of effect of 6-Aminonicotinamide on metabolism of RIF-1 tumor cells in vitro. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 8. pp. 4113-4119.
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