Comparison of fluorescence, (31)P NMR, and (7)Li NMR spectroscopic methods for investigating Li(+)/Mg(2+) competition for biomolecules

Anal Biochem. 1999 Jul 15;272(1):1-7. doi: 10.1006/abio.1999.4169.

Abstract

The biochemical action of lithium in the treatment of manic-depressive illness is still unknown. One hypothesis is that Li(+) competes for Mg(2+)-binding sites in biomolecules. We report here our studies on metal ion competition by three distinct methods: fluorescence, (31)P NMR, and (7)Li NMR spectroscopy, using ATP as a model ligand. By fluorescence spectroscopy, we used the dye, furaptra, by measuring the increases in Mg(2+) levels in an ATP solution as Li(+) levels were increased in the solution. This increase in Mg(2+) levels was indicated by increases in the fluorescence intensity ratio (335/370) of furaptra. By (31)P NMR spectroscopy, this competition was demonstrated by changes in the (31)P NMR spectrum of ATP. The Li(+)/Mg(2+) competition was indicated by predictable changes in the separation between the alpha and beta resonances of the phosphates of ATP. For (7)Li NMR spectroscopy, spin-lattice relaxation measurements were used, which provided free Li(+) concentrations that could be used for determining the free Mg(2+) values in ATP solutions. The values of the free Mg(2+) concentrations obtained by all three methods were in good agreement. The fluorescence and (7)Li NMR methods, however, proved to be more sensitive to low concentrations of Li(+) than the (31)P NMR method.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Binding Sites
  • Binding, Competitive
  • Bipolar Disorder / drug therapy
  • Bipolar Disorder / metabolism
  • Humans
  • Lithium / metabolism*
  • Magnesium / metabolism*
  • Magnetic Resonance Spectroscopy / methods*
  • Phosphates / metabolism
  • Phosphorus
  • Spectrometry, Fluorescence / methods*

Substances

  • Phosphates
  • Phosphorus
  • Adenosine Triphosphate
  • Lithium
  • Magnesium