N‐acetyl‐β‐D‐glucopyranosylamine

A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose

N. G. Oikonomakos, M. Kontou, S. E. Zographos, K. A. Watson, L. N. Johnson, C. J.F. Bichard, G. W.J. Fleet, K. R. Acharya

Research output: Contribution to journalArticle

Abstract

Structure‐based drug design has led to the discovery of a number of glucose analogue inhibitors of glycogen phosphorylase that have an increased affinity compared to α‐D‐glucose (Ki = 1.7 mM). The best inhibitor in the class of N‐acyl derivatives of β‐D‐glucopyranosylamine, N‐acetyl‐β‐D‐glucopyranosylamine (1‐GlcNAc), has been characterized by kinetic, ultracentrifugation, and crystallographic studies. 1‐GlcNAc acts as a competitive inhibitor for both the b (Ki = 32 μM) and the α (Ki = 35 μM) forms of the enzyme with respect to glucose 1‐phosphate and in synergism with caffeine, mimicking the binding of glucose. Sedimentation velocity experiments demonstrated that 1‐GlcNAc was able to induce dissociation of tetrameric phosphorylase α and stabilization of the dimeric T‐state conformation. Co‐crystals of the phosphorylase b‐1‐GlcNAc‐IMP complex were grown in space group P43212, with native‐like unit cell dimensions, and the complex structure has been refined to give a crystallographic R factor of 18.1%, for data between 8 and 2.3 Å resolution. 1‐GlcNAc binds tightly at the catalytic site of T‐state phosphorylase b at approximately the same position as that of α‐D‐glucose. The ligand can be accommodated in the catalytic site with very little change in the protein structure and stabilizes the T‐state conformation of the 280s loop by making several favorable contacts to Asn 284 of this loop. Structural comparisons show that the T‐state phosphorylase b‐1‐GlcNAc‐IMP complex structure is overall similar to the T‐state phosphorylase b‐α‐D‐glucose complex structure. The structure of the 1‐GlcNAc complex provides a rational for the biochemical properties of the inhibitor.

Original languageEnglish
Pages (from-to)2469-2477
Number of pages9
JournalProtein Science
Volume4
Issue number12
DOIs
Publication statusPublished - 1 Jan 1995

Keywords

  • binding
  • glycogen phosphorylase
  • inhibition
  • N‐acetyl‐β‐D‐gIucopyranosylamine

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

N‐acetyl‐β‐D‐glucopyranosylamine : A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose. / Oikonomakos, N. G.; Kontou, M.; Zographos, S. E.; Watson, K. A.; Johnson, L. N.; Bichard, C. J.F.; Fleet, G. W.J.; Acharya, K. R.

In: Protein Science, Vol. 4, No. 12, 01.01.1995, p. 2469-2477.

Research output: Contribution to journalArticle

Oikonomakos, NG, Kontou, M, Zographos, SE, Watson, KA, Johnson, LN, Bichard, CJF, Fleet, GWJ & Acharya, KR 1995, 'N‐acetyl‐β‐D‐glucopyranosylamine: A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose', Protein Science, vol. 4, no. 12, pp. 2469-2477. https://doi.org/10.1002/pro.5560041203
Oikonomakos, N. G. ; Kontou, M. ; Zographos, S. E. ; Watson, K. A. ; Johnson, L. N. ; Bichard, C. J.F. ; Fleet, G. W.J. ; Acharya, K. R. / N‐acetyl‐β‐D‐glucopyranosylamine : A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose. In: Protein Science. 1995 ; Vol. 4, No. 12. pp. 2469-2477.
@article{9ea2765c1a184db68cf031f91553a063,
title = "N‐acetyl‐β‐D‐glucopyranosylamine: A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose",
abstract = "Structure‐based drug design has led to the discovery of a number of glucose analogue inhibitors of glycogen phosphorylase that have an increased affinity compared to α‐D‐glucose (Ki = 1.7 mM). The best inhibitor in the class of N‐acyl derivatives of β‐D‐glucopyranosylamine, N‐acetyl‐β‐D‐glucopyranosylamine (1‐GlcNAc), has been characterized by kinetic, ultracentrifugation, and crystallographic studies. 1‐GlcNAc acts as a competitive inhibitor for both the b (Ki = 32 μM) and the α (Ki = 35 μM) forms of the enzyme with respect to glucose 1‐phosphate and in synergism with caffeine, mimicking the binding of glucose. Sedimentation velocity experiments demonstrated that 1‐GlcNAc was able to induce dissociation of tetrameric phosphorylase α and stabilization of the dimeric T‐state conformation. Co‐crystals of the phosphorylase b‐1‐GlcNAc‐IMP complex were grown in space group P43212, with native‐like unit cell dimensions, and the complex structure has been refined to give a crystallographic R factor of 18.1{\%}, for data between 8 and 2.3 {\AA} resolution. 1‐GlcNAc binds tightly at the catalytic site of T‐state phosphorylase b at approximately the same position as that of α‐D‐glucose. The ligand can be accommodated in the catalytic site with very little change in the protein structure and stabilizes the T‐state conformation of the 280s loop by making several favorable contacts to Asn 284 of this loop. Structural comparisons show that the T‐state phosphorylase b‐1‐GlcNAc‐IMP complex structure is overall similar to the T‐state phosphorylase b‐α‐D‐glucose complex structure. The structure of the 1‐GlcNAc complex provides a rational for the biochemical properties of the inhibitor.",
keywords = "binding, glycogen phosphorylase, inhibition, N‐acetyl‐β‐D‐gIucopyranosylamine",
author = "Oikonomakos, {N. G.} and M. Kontou and Zographos, {S. E.} and Watson, {K. A.} and Johnson, {L. N.} and Bichard, {C. J.F.} and Fleet, {G. W.J.} and Acharya, {K. R.}",
year = "1995",
month = "1",
day = "1",
doi = "10.1002/pro.5560041203",
language = "English",
volume = "4",
pages = "2469--2477",
journal = "Protein Science",
issn = "0961-8368",
publisher = "Cold Spring Harbor Laboratory Press",
number = "12",

}

TY - JOUR

T1 - N‐acetyl‐β‐D‐glucopyranosylamine

T2 - A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose

AU - Oikonomakos, N. G.

AU - Kontou, M.

AU - Zographos, S. E.

AU - Watson, K. A.

AU - Johnson, L. N.

AU - Bichard, C. J.F.

AU - Fleet, G. W.J.

AU - Acharya, K. R.

PY - 1995/1/1

Y1 - 1995/1/1

N2 - Structure‐based drug design has led to the discovery of a number of glucose analogue inhibitors of glycogen phosphorylase that have an increased affinity compared to α‐D‐glucose (Ki = 1.7 mM). The best inhibitor in the class of N‐acyl derivatives of β‐D‐glucopyranosylamine, N‐acetyl‐β‐D‐glucopyranosylamine (1‐GlcNAc), has been characterized by kinetic, ultracentrifugation, and crystallographic studies. 1‐GlcNAc acts as a competitive inhibitor for both the b (Ki = 32 μM) and the α (Ki = 35 μM) forms of the enzyme with respect to glucose 1‐phosphate and in synergism with caffeine, mimicking the binding of glucose. Sedimentation velocity experiments demonstrated that 1‐GlcNAc was able to induce dissociation of tetrameric phosphorylase α and stabilization of the dimeric T‐state conformation. Co‐crystals of the phosphorylase b‐1‐GlcNAc‐IMP complex were grown in space group P43212, with native‐like unit cell dimensions, and the complex structure has been refined to give a crystallographic R factor of 18.1%, for data between 8 and 2.3 Å resolution. 1‐GlcNAc binds tightly at the catalytic site of T‐state phosphorylase b at approximately the same position as that of α‐D‐glucose. The ligand can be accommodated in the catalytic site with very little change in the protein structure and stabilizes the T‐state conformation of the 280s loop by making several favorable contacts to Asn 284 of this loop. Structural comparisons show that the T‐state phosphorylase b‐1‐GlcNAc‐IMP complex structure is overall similar to the T‐state phosphorylase b‐α‐D‐glucose complex structure. The structure of the 1‐GlcNAc complex provides a rational for the biochemical properties of the inhibitor.

AB - Structure‐based drug design has led to the discovery of a number of glucose analogue inhibitors of glycogen phosphorylase that have an increased affinity compared to α‐D‐glucose (Ki = 1.7 mM). The best inhibitor in the class of N‐acyl derivatives of β‐D‐glucopyranosylamine, N‐acetyl‐β‐D‐glucopyranosylamine (1‐GlcNAc), has been characterized by kinetic, ultracentrifugation, and crystallographic studies. 1‐GlcNAc acts as a competitive inhibitor for both the b (Ki = 32 μM) and the α (Ki = 35 μM) forms of the enzyme with respect to glucose 1‐phosphate and in synergism with caffeine, mimicking the binding of glucose. Sedimentation velocity experiments demonstrated that 1‐GlcNAc was able to induce dissociation of tetrameric phosphorylase α and stabilization of the dimeric T‐state conformation. Co‐crystals of the phosphorylase b‐1‐GlcNAc‐IMP complex were grown in space group P43212, with native‐like unit cell dimensions, and the complex structure has been refined to give a crystallographic R factor of 18.1%, for data between 8 and 2.3 Å resolution. 1‐GlcNAc binds tightly at the catalytic site of T‐state phosphorylase b at approximately the same position as that of α‐D‐glucose. The ligand can be accommodated in the catalytic site with very little change in the protein structure and stabilizes the T‐state conformation of the 280s loop by making several favorable contacts to Asn 284 of this loop. Structural comparisons show that the T‐state phosphorylase b‐1‐GlcNAc‐IMP complex structure is overall similar to the T‐state phosphorylase b‐α‐D‐glucose complex structure. The structure of the 1‐GlcNAc complex provides a rational for the biochemical properties of the inhibitor.

KW - binding

KW - glycogen phosphorylase

KW - inhibition

KW - N‐acetyl‐β‐D‐gIucopyranosylamine

UR - http://www.scopus.com/inward/record.url?scp=0028972335&partnerID=8YFLogxK

U2 - 10.1002/pro.5560041203

DO - 10.1002/pro.5560041203

M3 - Article

VL - 4

SP - 2469

EP - 2477

JO - Protein Science

JF - Protein Science

SN - 0961-8368

IS - 12

ER -