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Mapping the Binding Site of an Aptamer on ATP Using MicroScale Thermophoresis
Published on: January 7, 2017
DNA aptamer-mediated regulation of the hairpin ribozyme by human alpha-thrombin
S Hani Najafi-Shoushtari1, Michael Famulok
1LIMES Program Unit Chemical Biology and Medicinal Chemistry, c/o Kekulé Institut für Organische Chemie und Biochemie, University of Bonn, Gerhard-Domagk-Strasse 1, 53121 Bonn, Germany.
Abstract:
The combination of specific ligand-binding aptamers with hairpin ribozyme catalysis generates molecules that can be controlled by external factors. Here we have generated hairpin ribozymes that can be regulated by a short DNA aptamer specific for human alpha-thrombin. This was achieved by constructing a ribozyme variant harboring an RNA sequence complementary to the aptamer, to which the aptamer can hybridize forming a heteroduplex. In this way, the DNA aptamer completely abolishes the catalytic activity of the ribozyme, due to the formation of an inactive ribozyme conformation. However, in the presence of the aptamer's target protein human alpha-thrombin, the inhibitory effect of the DNA aptamer is competitively neutralized and the ribozyme is activated in a highly specific fashion. Protein-responsive allosteric ribozymes are proposed to act as tools with potential applications in medicine where fast detection of clinically relevant targets is required.
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