Interaction of human 3-phosphoglycerate kinase with l-ADP, the mirror image of d-ADP

A. Varga, Judit Szabó, Beáta Flachner, Béatrice Roy, Peter Konarev, Dmitri Svergun, P. Závodszky, Christian Périgaud, Tom Barman, Corinne Lionne, M. Vas

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

l-Nucleoside-analogues, mirror images of the natural d-nucleosides, are a new class of antiviral and anticancer agents. In the cell they have to be phosphorylated to pharmacologically active triphosphate forms, the last step seems to involve human 3-phosphoglycerate kinase (hPGK). Here we present a steady state kinetic and biophysical study of the interaction of the model compound l-MgADP with hPGK. l-MgADP is a good substrate with kcat and Km values of 685 s-1 and 0.27 mM, respectively. Double inhibition studies suggest that l-MgADP binds to the specific adenosine-binding site and protects the conformation of hPGK molecule against heat denaturation, as detected by microcalorimetry. Structural details of the interaction in the enzyme active site are different for the d- and l-enantiomers (e.g. the effect of Mg2+), but these differences do not prevent the occurrence of the catalytic cycle, which is accompanied by the hinge-bending domain closure, as indicated by SAXS measurements.

Original languageEnglish
Pages (from-to)994-1000
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume366
Issue number4
DOIs
Publication statusPublished - Feb 22 2008

Fingerprint

Phosphoglycerate Kinase
Adenosine Diphosphate
Mirrors
Nucleosides
Denaturation
Enantiomers
Hinges
Antineoplastic Agents
Adenosine
Antiviral Agents
Conformations
Catalytic Domain
Hot Temperature
Binding Sites
Molecules
Kinetics
Substrates
Enzymes
3-phosphoglycerate

Keywords

  • Enantioselectivity
  • Enzyme kinetic analysis
  • Human 3-phosphoglycerate kinase
  • Microcalorimetry
  • Nucleotide substrates
  • Small angle X-ray scattering

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Interaction of human 3-phosphoglycerate kinase with l-ADP, the mirror image of d-ADP. / Varga, A.; Szabó, Judit; Flachner, Beáta; Roy, Béatrice; Konarev, Peter; Svergun, Dmitri; Závodszky, P.; Périgaud, Christian; Barman, Tom; Lionne, Corinne; Vas, M.

In: Biochemical and Biophysical Research Communications, Vol. 366, No. 4, 22.02.2008, p. 994-1000.

Research output: Contribution to journalArticle

Varga, A. ; Szabó, Judit ; Flachner, Beáta ; Roy, Béatrice ; Konarev, Peter ; Svergun, Dmitri ; Závodszky, P. ; Périgaud, Christian ; Barman, Tom ; Lionne, Corinne ; Vas, M. / Interaction of human 3-phosphoglycerate kinase with l-ADP, the mirror image of d-ADP. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 366, No. 4. pp. 994-1000.
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