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. Author manuscript; available in PMC: 2013 Jan 11.
Published in final edited form as: J Am Chem Soc. 2011 Dec 14;134(1):256–262. doi: 10.1021/ja206455t

Figure 1.

Figure 1

Pt(II)-induced crosslinking of the Hammerhead ribozyme. a) The sequence of HHRzES-SS(dC17,C1.1ps) ribozyme used in this study (left) consists of an ’enzyme strand’ (ES, blue) and phosphorothioate substituted ’substrate strand’ (SS, black).’PS’ (red) denotes the phosphorothioate substitution installed at the HHRz cleavage site and U-loop depicts a non-folding mutant. On the right, a denaturing gel depicting crosslinks formed by modified HHrz components. Gel lanes from left to right (− or + platinum): (i) isolated PS-modified SS (ii) isolated HHRz enzyme strand, (iii) the PSsubstituted HHRzES-SS ribozyme, (iv) a non-PS-substituted HHrzES-SS ribozyme, (v) a PS-substituted HHHrzES-SS ribozyme with a folding-deficient U-loop mutation, (vi) a repeat experiment using the PS-substitute HHRzES-SS ribozyme. Conditions (see Methods): 40 μM RNA in 1mM Mg(NO3)2, 100 mM NaNO3, 10 mM Na2PO4, pH 7.0, 16 h, 37°C, ± 3 equivalents activated [cis-diammine Pt(II)] analyzed by 20% dPAGE and stained with methylene blue.