Binding of ATP to the fructose-2,6-bisphosphatase domain of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase leads to activation of its 6-phosphofructo-2-kinase

The Journal of Biological Chemistry
Q H YangL Li

Abstract

To understand the mechanism by which the activity of the 6-phosphofructo-2-kinase (6PF-2K) of chicken liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase is stimulated by its substrate ATP, we studied two mutants of the enzyme. Mutation of either Arg-279, the penultimate basic residue within the Walker A nucleotide-binding fold in the bisphosphatase domain, or Arg-359 to Ala eliminated the activation of the chicken 6PF-2K by ATP. Binding analysis by fluorescence spectroscopy using 2'(3')-O-(N-methylanthraniloyl)-ATP revealed that the kinase domains of these two mutants, unlike that of the wild type enzyme, showed no cooperativity in ATP binding and that the mutant enzymes possess only the high affinity ATP binding site, suggesting that the ATP binding site on the bisphosphatase domain represents the low affinity site. This conclusion was supported by the result that the affinity of ATP for the isolated bisphosphatase domain is similar to that for the low affinity site in the wild type enzyme. In addition, we found that the 6PF-2K of a chimeric enzyme, in which the last 25 residues of chicken enzyme were replaced with those of the rat enzyme, could not be activated by ATP, despite the fact that the ATP-binding properties ...Continue Reading

References

Dec 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·J F BazanS J Pilkis
Jul 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·M R el-MaghrabiS J Pilkis
May 1, 1982·European Journal of Biochemistry·E van SchaftingenH G Hers
Jun 1, 1995·Protein Science : a Publication of the Protein Society·I J Kurland, S J Pilkis
Jan 1, 1995·Annual Review of Biochemistry·S J PilkisA J Lange
Apr 26, 1995·Biochemical and Biophysical Research Communications·L LiG J Xu
Jan 29, 1993·Biochemical and Biophysical Research Communications·L LiS J Pilkis
Jan 1, 1993·Progress in Nucleic Acid Research and Molecular Biology·G G Rousseau, L Hue

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Jul 24, 2007·Journal of Molecular Graphics & Modelling·M S ShaikhP V Bharatam

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