The novel fluorescent CDP-analogue (Pbeta)MABA-CDP is a specific probe for the NMP binding site of UMP/CMP kinase

Protein Sci. 1999 Dec;8(12):2697-704. doi: 10.1110/ps.8.12.2697.

Abstract

Direct thermodynamic and kinetic investigations of the binding of nucleotides to the nucleoside monophosphate (NMP) site of NMP kinases have not been possible so far because a spectroscopic probe was not available. By coupling a fluorescent N-methylanthraniloyl- (mant) group to the beta-phosphate of CDP via a butyl linker, a CDP analogue [(Pbeta)MABA-CDP] was obtained that still binds specifically to the NMP site of UmpKdicty, because the base and the ribose moieties, which are involved in specific interactions, are not modified. This allows the direct determination of binding constants for its substrates in competition experiments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / chemistry
  • Animals
  • Binding Sites
  • Cytidine Diphosphate / analogs & derivatives*
  • Cytidine Diphosphate / chemical synthesis
  • Cytidine Diphosphate / chemistry*
  • Dictyostelium / chemistry*
  • Fluorescent Dyes / chemical synthesis
  • Fluorescent Dyes / chemistry*
  • Kinetics
  • Magnetic Resonance Spectroscopy
  • Nucleoside-Phosphate Kinase / chemistry*
  • Pyrimidinones / chemistry*
  • Spectrometry, Fluorescence

Substances

  • Fluorescent Dyes
  • Pyrimidinones
  • cytidinediphospho-beta-(N'-methylanthraniloylaminobutyl)phosphoramidate
  • Cytidine Diphosphate
  • Adenosine Triphosphate
  • uridine monophosphate kinase
  • Nucleoside-Phosphate Kinase