The binding of 2-deoxy-o-glucose 6-phosphate to glycogen phosphorylase b

Kinetic and crystallographic studies

N. G. Oikonomakos, S. E. Zographos, L. N. Johnson, A. C. Papageorgiou, K. R. Acharya

Research output: Contribution to journalArticle

Abstract

Kinetic and crystallographic studies have characterized the effect of 2-deoxy-glucose 6-phosphate on the catalytic and structural properties of glycogen phosphorylase b. Previous work on the binding of glucose 6-phosphate, a potent physiological inhibitor of the enzyme, to T state phosphorylase b in the crystal showed that the inhibitor binds at the allosteric site and induces substantial conformational changes that affect the subunit-subunit interface. The hydrogen-bond from the O-2 hydroxyl of glucose 6-phosphate to the main-chain oxygen of Val40′ represents the only hydrogen bond from the sugar to the other subunit, and this interaction appears important for promoting a more "tensed" structure than native T state phosphorylase b. 2-Deoxy-glucose 6-phosphate acts competitively with both the activator AMP and the substrate glucose 1-phosphate, with Ki values of 0.53 mM and 1.23 mM, respectively. The binding of 2-deoxy-glucose 6-phosphate to T state glycogen phosphorylase b in the crystal, has been investigated and the complex phosphorylase b: 2-deoxy-glucose 6-phosphate has been refined to give a crystallographic R factor of 17.3%, for data between 8 Å and 2.3 Å. 2-Deoxy-glucose 6-phosphate binds at the allosteric site as the a anomer and adopts a different conformation compared to glucose 6-phosphate. The two conformations differ by 160° in the torsion angle about the C-5-C-6 bond. The contacts from the phosphate group are essentially identical to those made by the phosphate of glucose 6-phosphate but the 2-deoxy glucosyl moiety binds in a quite different orientation compared to the glucosyl of glucose 6-phosphate. 2-Deoxy-glucose 6-phosphate can be accommodated in the allosteric site with very little change in the protein, while structural comparisons show that the phosphorylase b: 2-deoxy-glucose 6-phosphate complex structure is overall more similar to a glucose-like complex than to the Glc-6-P complex structure.

Original languageEnglish
Pages (from-to)900-917
Number of pages18
JournalJournal of Molecular Biology
Volume254
Issue number5
DOIs
Publication statusPublished - 1 Jan 1995

Keywords

  • 2-deoxy-glucose 6-phosphate
  • Allosteric inhibition
  • Conformational changes
  • Glucose 6-phosphate
  • Glycogen phosphorylase

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

The binding of 2-deoxy-o-glucose 6-phosphate to glycogen phosphorylase b : Kinetic and crystallographic studies. / Oikonomakos, N. G.; Zographos, S. E.; Johnson, L. N.; Papageorgiou, A. C.; Acharya, K. R.

In: Journal of Molecular Biology, Vol. 254, No. 5, 01.01.1995, p. 900-917.

Research output: Contribution to journalArticle

Oikonomakos, N. G. ; Zographos, S. E. ; Johnson, L. N. ; Papageorgiou, A. C. ; Acharya, K. R. / The binding of 2-deoxy-o-glucose 6-phosphate to glycogen phosphorylase b : Kinetic and crystallographic studies. In: Journal of Molecular Biology. 1995 ; Vol. 254, No. 5. pp. 900-917.
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AB - Kinetic and crystallographic studies have characterized the effect of 2-deoxy-glucose 6-phosphate on the catalytic and structural properties of glycogen phosphorylase b. Previous work on the binding of glucose 6-phosphate, a potent physiological inhibitor of the enzyme, to T state phosphorylase b in the crystal showed that the inhibitor binds at the allosteric site and induces substantial conformational changes that affect the subunit-subunit interface. The hydrogen-bond from the O-2 hydroxyl of glucose 6-phosphate to the main-chain oxygen of Val40′ represents the only hydrogen bond from the sugar to the other subunit, and this interaction appears important for promoting a more "tensed" structure than native T state phosphorylase b. 2-Deoxy-glucose 6-phosphate acts competitively with both the activator AMP and the substrate glucose 1-phosphate, with Ki values of 0.53 mM and 1.23 mM, respectively. The binding of 2-deoxy-glucose 6-phosphate to T state glycogen phosphorylase b in the crystal, has been investigated and the complex phosphorylase b: 2-deoxy-glucose 6-phosphate has been refined to give a crystallographic R factor of 17.3%, for data between 8 Å and 2.3 Å. 2-Deoxy-glucose 6-phosphate binds at the allosteric site as the a anomer and adopts a different conformation compared to glucose 6-phosphate. The two conformations differ by 160° in the torsion angle about the C-5-C-6 bond. The contacts from the phosphate group are essentially identical to those made by the phosphate of glucose 6-phosphate but the 2-deoxy glucosyl moiety binds in a quite different orientation compared to the glucosyl of glucose 6-phosphate. 2-Deoxy-glucose 6-phosphate can be accommodated in the allosteric site with very little change in the protein, while structural comparisons show that the phosphorylase b: 2-deoxy-glucose 6-phosphate complex structure is overall more similar to a glucose-like complex than to the Glc-6-P complex structure.

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