The binding of ATP and AMP to Escherichia coli adenylate kinase investigated by 1H and 15N NMR spectroscopy

John Glushka, Octavian Bârzu, Robert S. Sarfati, Vinod K. Kansal, David Cowburn

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

[N6 15N]ATP and [N6 15N]AMP, complexed with E.coli adenylate kinase (AKe), were observed with 15N isotope-filtered NMR pulse sequences and 1H{15N} heterocorrelated experiments to determine differences between binding sites based on chemical shifts and competition by substrate analogs. The chemical shifts of the N6 amino proton and nitrogen signals changed significantly after mixing with adenylate kinase. Differences in chemical shifts between the bound ATP and AMP signals are slight. The response of these shifts to further addition of other substrates or Mg2+ supports the view that the unchelated nucleotides can bind to both the sites, whereas the metal complexed species are restricted to the MgATP MgADP binding site.

Original languageEnglish (US)
Pages (from-to)432-438
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume172
Issue number2
DOIs
StatePublished - Oct 30 1990
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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