Atomic details of near-transition state conformers for enzyme phosphoryl transfer revealed by MgF3 - rather than by phosphoranes

Nicola J. Baxter, Matthew W. Bowler, Tooba Alizadeh, Matthew J. Cliff, Andrea M. Hounslow, Bin Wu, David B Berkowitz, Nicholas H. Williams, G. Michael Blackburn, Jonathan P. Waltho

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Abstract

Prior evidence supporting the direct observation of phosphorane intermediates in enzymatic phosphoryl transfer reactions was based on the interpretation of electron density corresponding to trigonal species bridging the donor and acceptor atoms. Close examination of the crystalline state of β-phosphoglucomutase, the archetypal phosphorane intermediate-containing enzyme, reveals that the trigonal species is not PO3 - , but is MgF3 - (trifluoromagnesate). Although MgF 3 - complexes are transition state analogues rather than phosphoryl group transfer reaction intermediates, the presence of fluorine nuclei in near-transition state conformations offers new opportunities to explore the nature of the interactions, in particular the independent measures of local electrostatic and hydrogen-bonding distributions using 19F NMR. Measurements on three β-PGM-MgF3 - -sugar phosphate complexes show a remarkable relationship between NMR chemical shifts, primary isotope shifts, NOEs, cross hydrogen bond F···H-N scalar couplings, and the atomic positions determined from the high-resolution crystal structure of the β-PGM-MgF3 - -G6P complex. The measurements provide independent validation of the structural and isoelectronic MgF3 - model of near-transition state conformations.

Original languageEnglish (US)
Pages (from-to)4555-4560
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number10
DOIs
StatePublished - Mar 9 2010

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Phosphoranes
Sugar Phosphates
Phosphoglucomutase
Fluorine
Enzymes
Hydrogen Bonding
Static Electricity
Isotopes
Hydrogen
Observation
Electrons

Keywords

  • 19F NMR
  • Phosphoryl transfer enzyme
  • Transition state analogue
  • Trifluoromagnesate

ASJC Scopus subject areas

  • General

Cite this

Atomic details of near-transition state conformers for enzyme phosphoryl transfer revealed by MgF3 - rather than by phosphoranes. / Baxter, Nicola J.; Bowler, Matthew W.; Alizadeh, Tooba; Cliff, Matthew J.; Hounslow, Andrea M.; Wu, Bin; Berkowitz, David B; Williams, Nicholas H.; Blackburn, G. Michael; Waltho, Jonathan P.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, No. 10, 09.03.2010, p. 4555-4560.

Research output: Contribution to journalArticle

Baxter, Nicola J. ; Bowler, Matthew W. ; Alizadeh, Tooba ; Cliff, Matthew J. ; Hounslow, Andrea M. ; Wu, Bin ; Berkowitz, David B ; Williams, Nicholas H. ; Blackburn, G. Michael ; Waltho, Jonathan P. / Atomic details of near-transition state conformers for enzyme phosphoryl transfer revealed by MgF3 - rather than by phosphoranes. In: Proceedings of the National Academy of Sciences of the United States of America. 2010 ; Vol. 107, No. 10. pp. 4555-4560.
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abstract = "Prior evidence supporting the direct observation of phosphorane intermediates in enzymatic phosphoryl transfer reactions was based on the interpretation of electron density corresponding to trigonal species bridging the donor and acceptor atoms. Close examination of the crystalline state of β-phosphoglucomutase, the archetypal phosphorane intermediate-containing enzyme, reveals that the trigonal species is not PO3 - , but is MgF3 - (trifluoromagnesate). Although MgF 3 - complexes are transition state analogues rather than phosphoryl group transfer reaction intermediates, the presence of fluorine nuclei in near-transition state conformations offers new opportunities to explore the nature of the interactions, in particular the independent measures of local electrostatic and hydrogen-bonding distributions using 19F NMR. Measurements on three β-PGM-MgF3 - -sugar phosphate complexes show a remarkable relationship between NMR chemical shifts, primary isotope shifts, NOEs, cross hydrogen bond F···H-N scalar couplings, and the atomic positions determined from the high-resolution crystal structure of the β-PGM-MgF3 - -G6P complex. The measurements provide independent validation of the structural and isoelectronic MgF3 - model of near-transition state conformations.",
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AB - Prior evidence supporting the direct observation of phosphorane intermediates in enzymatic phosphoryl transfer reactions was based on the interpretation of electron density corresponding to trigonal species bridging the donor and acceptor atoms. Close examination of the crystalline state of β-phosphoglucomutase, the archetypal phosphorane intermediate-containing enzyme, reveals that the trigonal species is not PO3 - , but is MgF3 - (trifluoromagnesate). Although MgF 3 - complexes are transition state analogues rather than phosphoryl group transfer reaction intermediates, the presence of fluorine nuclei in near-transition state conformations offers new opportunities to explore the nature of the interactions, in particular the independent measures of local electrostatic and hydrogen-bonding distributions using 19F NMR. Measurements on three β-PGM-MgF3 - -sugar phosphate complexes show a remarkable relationship between NMR chemical shifts, primary isotope shifts, NOEs, cross hydrogen bond F···H-N scalar couplings, and the atomic positions determined from the high-resolution crystal structure of the β-PGM-MgF3 - -G6P complex. The measurements provide independent validation of the structural and isoelectronic MgF3 - model of near-transition state conformations.

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