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. 1978 Sep;75(9):4204–4208. doi: 10.1073/pnas.75.9.4204

Spectroscopic sensitivity of linear-benzoadenine nucleotides to divalent metal counterions, side chain conformations, micelles, and enzymes

Pieter VanDerLijn 1, Jorge R Barrio 1, Nelson J Leonard 1,*
PMCID: PMC336080  PMID: 16592561

Abstract

From pKa data for lin-benzoadenosine 5′-mono-, 5′-di-, and 5′-triphosphates, which are fluorescent “stretched-out” analogues of adenine nucleotides, it was possible to designate the cases of interaction of phosphate with the heteroaromatic moiety. The addition of divalent metal cations or quaternary ammonium micelles diminishes the direct intramolecular interaction between the phosphate(s) and base and consequently brings the pKa values close to that of lin-benzoadenosine. Fluorescence spectroscopy was used to investigate the interaction of lin-benzoadenine nucleotides with Mg2+, Mn2+, and Co2+. The association constants for the formation of such complexes were obtained from measurements of steady-state fluorescence quenching. Phase and modulation measurements of the fluorescence lifetimes of lin-benzoadenine nucleotides as a function of Co2+ concentration permitted determination of the static component of the quenching due to intramolecular complex formation. The association constants of the lin-benzoadenine nucleotides with all of the divalent metal ions studied were greater than those observed for the corresponding adenine nucleotides and were in the order: lin-benzo-ATP > lin-benzo-ADP > lin-benzo-AMP. Fourier transform 1H NMR of lin-benzo-ATP in the presence of Co2+ showed broadening of the aromatic proton signals, the 2-H signal (corresponding to the 8-H in ATP) being the most affected. Models are proposed to explain the phosphate-base interaction, the influence of metal ions on base protonation, and the intramo- the intramolecular quenching observed in the complexes due to paramagnetic ion (Co2+, Mn2+) and base interaction.

Keywords: adenine binding sites, dimensional probes, fluorescence, phosphate-adenine interaction, pKa

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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