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• W2557993811 abstract "Abstract Mature erythrocytes are unable to synthesize AMP from IMP due to the developmental loss of adenylosuccinate synthetase, which also effectively prevents the synthesis of adenine nucleotides from circulating hypoxanthine, the major salvageable purine compound. Consequently, AMP deaminase (AMPD) activity must be tightly regulated in order to avoid irreversible depletion of the erythrocyte adenine nucleotide pool during periods of energy imbalance. However, increased intracellular calcium promotes activation of erythrocyte AMPD (AMPD3) through a protein-protein interaction with calmodulin (Mahnke and Sabina, Biochemistry44:5551, 2005). Therefore, we hypothesized that Sickle Cell Disease (SCD) erythrocytes would more rapidly accumulate IMP and deplete the adenine nucleotide pool during periods of energy imbalance. In order to test this hypothesis, acid-soluble pools of these metabolites were quantified by anion-exchange HPLC in SCD and control erythrocytes during autoincubation and glucose starvation, both at 37C. Data in the table show that SCD erythrocytes accumulate more IMP during a 6-hour autoincubation compared to control cells, and this is accompanied by a decrease in AMP. Effect of Autoincubation on Erythrocyte AMP and IMP Pools GROUP HOURS AMPa IMP IMP/AMP a, metabolite concentrations reported as μmol/g Hb; †, p<.05 when compared to 0 time in a paired student’s t-test; ‡, p<.05 when compared to control in an unpaired student’s t-test Control (N=4) 0 .29±.08 .15±.06 .58±.28 6 .29±.01 .33±.11† 1.15±.35† SCD (N=5) 0 .22±.04 .08±.03 .36±.10 6 .14±.03† .74±.24†‡ 5.15±1.25†‡ Data in the figure show an accelerated depletion of ATP and increased accumulation of IMP in glucose-starved SCD erythrocytes compared to control cells (*, p<.05 when compared to control in an unpaired student’s t-test). Figure Figure These combined results demonstrate AMPD3 enzyme activation and accelerated adenine nucleotide pool depletion during periods of energy imbalance in SCD erythrocytes. This metabolic imbalance in SCD erythrocytes could contribute to 1) an increased rate of hemolysis, 2) cation channel dysregulation, 3) modulation of adhesive membrane components, and 4) diminished protein phosphorylation. Furthermore, strategies directed against this metabolic imbalance, e.g., diffusible AMPD inhibitors and adenine supplementation, may have a therapeutic benefit in SCD." @default.
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• W2557993811 date "2005-11-16" @default.
• W2557993811 modified "2023-09-30" @default.
• W2557993811 title "AMP Deaminase Activation Contributes to Accelerated Adenine Nucleotide Pool Depletion during Periods of Energy Imbalance in Sickle Cell Erythrocytes." @default.
• W2557993811 doi "https://doi.org/10.1182/blood.v106.11.1671.1671" @default.
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