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. 1999 Aug;5(8):1099–1104. doi: 10.1017/s1355838299990738

Identification and characterization of a novel high affinity metal-binding site in the hammerhead ribozyme.

M R Hansen 1, J P Simorre 1, P Hanson 1, V Mokler 1, L Bellon 1, L Beigelman 1, A Pardi 1
PMCID: PMC1369832  PMID: 10445883

Abstract

A novel metal-binding site has been identified in the hammerhead ribozyme by 31P NMR. The metal-binding site is associated with the A13 phosphate in the catalytic core of the hammerhead ribozyme and is distinct from any previously identified metal-binding sites. 31P NMR spectroscopy was used to measure the metal-binding affinity for this site and leads to an apparent dissociation constant of 250-570 microM at 25 degrees C for binding of a single Mg2+ ion. The NMR data also show evidence of a structural change at this site upon metal binding and these results are compared with previous data on metal-induced structural changes in the core of the hammerhead ribozyme. These NMR data were combined with the X-ray structure of the hammerhead ribozyme (Pley HW, Flaherty KM, McKay DB. 1994. Nature 372:68-74) to model RNA ligands involved in binding the metal at this A13 site. In this model, the A13 metal-binding site is structurally similar to the previously identified A(g) metal-binding site and illustrates the symmetrical nature of the tandem G x A base pairs in domain 2 of the hammerhead ribozyme. These results demonstrate that 31P NMR represents an important method for both identification and characterization of metal-binding sites in nucleic acids.

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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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