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Quantitative Analysis of Purine Nucleotides Indicates That Purinosomes Increase de Novo Purine Biosynthesis
- Source :
- Journal of Biological Chemistry. 290:6705-6713
- Publication Year :
- 2015
- Publisher :
- Elsevier BV, 2015.
-
Abstract
- Enzymes in the de novo purine biosynthesis pathway are recruited to form a dynamic metabolic complex referred to as the purinosome. Previous studies have demonstrated that purinosome assembly responds to purine levels in culture medium. Purine-depleted medium or 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT) treatment stimulates the purinosome assembly in HeLa cells. Here, several metabolomic technologies were applied to quantify the static cellular levels of purine nucleotides and measure the de novo biosynthesis rate of IMP, AMP, and GMP. Direct comparison of purinosome-rich cells (cultured in purine-depleted medium) and normal cells showed a 3-fold increase in IMP concentration in purinosome-rich cells and similar levels of AMP, GMP, and ratios of AMP/GMP and ATP/ADP for both. In addition, a higher level of IMP was also observed in HeLa cells treated with DMAT. Furthermore, increases in the de novo IMP/AMP/GMP biosynthetic flux rate under purine-depleted condition were observed. The synthetic enzymes, adenylosuccinate synthase (ADSS) and inosine monophosphate dehydrogenase (IMPDH), downstream of IMP were also shown to be part of the purinosome. Collectively, these results provide further evidence that purinosome assembly is directly related to activated de novo purine biosynthesis, consistent with the functionality of the purinosome.
- Subjects :
- Purine
Spectrometry, Mass, Electrospray Ionization
Magnetic Resonance Spectroscopy
Biochemistry
Adenylosuccinate Synthase
chemistry.chemical_compound
IMP Dehydrogenase
Biosynthesis
Multienzyme Complexes
IMP dehydrogenase
Humans
Metabolomics
Nucleotide
skin and connective tissue diseases
Adenylosuccinate lyase
Purine metabolism
Purine Nucleotides
Molecular Biology
chemistry.chemical_classification
biology
Purinosome
Adenylosuccinate synthase
Cell Biology
Molecular biology
Metabolism
chemistry
Purines
biology.protein
Benzimidazoles
sense organs
Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor
HeLa Cells
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 290
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....f1022266b6bdc4408a852bff70af6517