Pb EXAFS Studies on DNA Quadruplexes: Identification of Metal Ion Binding Site
| Autor: | Richard H. Shafer, Jeffery T. Davis, Ivan Smirnov, Ingrid J. Pickering, Frank W. Kotch |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Circular dichroism
Binding Sites Guanosine Extended X-ray absorption fine structure Absorption spectroscopy Guanine Circular Dichroism Metal ions in aqueous solution Metal Binding Site DNA Crystallography X-Ray Biochemistry Metal Crystallography chemistry.chemical_compound Lead chemistry visual_art visual_art.visual_art_medium Guanine nucleoside |
| Zdroj: | Biochemistry. 41:12133-12139 |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | Nucleic acid quadruplexes are composed of guanine quartets stabilized by specific metal ions. X-ray diffraction can provide high-resolution information on the structure and metal binding properties of quadruplexes, but only if they can be crystallized. NMR can provide detailed information on the solution structure of such quadruplexes but little quantitative data concerning the metal binding site. Here we apply extended X-ray absorption fine structure (EXAFS) measurements to characterize the metal ion binding site, in frozen solution, of the unimolecular quadruplex formed by the thrombin binding aptamer, d(G(2)T(2)G(2)TGTG(2)T(2)G(2)) (TBA), in the presence of Pb(2+) ions. The Pb L(III) -edge X-ray absorption spectrum of this metal-DNA complex is very similar to that we obtain for a Pb(2+)-stabilized quartet system of known structure constructed from a modified guanine nucleoside (G1). The Fourier transforms of the Pb(2+) complexes with both TBA and G1 show a first-shell interaction at about 2.6 A, and a weaker, broader shell at 3.5-4.0 A. Quantitative analysis of the EXAFS data reveals the following: (i) very close agreement between interatomic distances at the metal coordination site for the Pb(2+)-G1 complex determined by EXAFS and by X-ray crystallography; (ii) similarly close agreement between interatomic distances measured by EXAFS for the Pb(2+)-G1 and Pb(2+)-TBA complexes. These results provide strong evidence for binding of the Pb(2+) ion in the region between the two quartets in the Pb(2+)-TBA complex, coordinated to the eight surrounding guanine O6 atoms. The specific binding of Pb(2+) to DNA examined here may be relevant to the genotoxic effects of this environmentally important heavy metal. Furthermore, these results demonstrate the utility of EXAFS as a method for quantitative characterization of specific metal binding sites in nucleic acids in solution. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|