Competition between Co(NH3)63+ and inner sphere Mg2+ ions in the HDV ribozyme

Bo Gong, Jui Hui Chen, Philip C. Bevilacqua, Barbara L. Golden, Paul R. Carey

Research output: Contribution to journalArticlepeer-review

32 Scopus citations


Divalent cations play critical structural and functional roles in many RNAs. While the hepatitis delta virus (HDV) ribozyme can undergo self-cleavage in the presence of molar concentrations of monovalent cations, divalent cations such as Mg2+ are required for efficient catalysis under physiological conditions. Moreover, the cleavage reaction can be inhibited with Co(NH 3)63+, an analogue of Mg(H2O) 62+. Here, the binding of Mg2+ and Co(NH 3)63+ to the HDV ribozyme is studied by Raman microscopic analysis of crystals. Raman difference spectra acquired at different metal ion conditions reveal changes in the ribozyme. When Mg2+ alone is introduced to the ribozyme, inner sphere coordination of Mg(H 2O)x2+ (x ≤ 5) to nonbridging PO 2- oxygen and changes in base stretches and phosphodiester group conformation are observed. In addition, binding of Mg2+ induces deprotonation of a cytosine assigned to the general acid C75, consistent with solution studies. When Co(NH3)63+ alone is introduced, deprotonation of C75 is again observed, as are distinctive changes in base vibrational ring modes and phosphodiester backbone conformation. In contrast to Mg2+ binding, Co(NH3)6 3+ binding does not perturb PO2- group vibrations, consistent with its ability to make only outer sphere contacts. Surprisingly, competitive binding studies reveal that Co(NH3) 63+ ions displace some inner sphere-coordinated magnesium species, including ions coordinated to PO2- groups or the N7 of a guanine, likely G1 at the active site. These observations contrast with the tenet that Co(NH3)63+ ions displace only outer sphere magnesium ions. Overall, our data support two classes of inner sphere Mg2+-PO2- binding sites: sites that Co(NH3)63+ can displace and others it cannot.

Original languageEnglish (US)
Pages (from-to)11961-11970
Number of pages10
Issue number50
StatePublished - Dec 22 2009

All Science Journal Classification (ASJC) codes

  • Biochemistry


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