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Selection of 2′-Fluoro-modified RNA Aptamers for Alleviation of Cocaine and MK-801Inhibition of the Nicotinic Acetylcholine Receptor

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Abstract

The nicotinic acetylcholine receptor (nAChR) belongs to a group of five stracturally related proteins that regulate signal transmission between approximately 1012 cells of the mammalian nervous system. Many therapeutic agents and abused drugs inhibit the nAChR, including the anti-convulsant MK-801 and the abused drug cocaine. Many attempts have been made to find compounds that prevent inhibition by cocaine. Use of transient kinetic techniques to investigate the inhibition of the receptor by MK-801 and cocaine led to an inhibition mechanism not previously proposed. The mechanism led to the development of combinatorially synthesized RNA ligands that alleviate inhibition of the receptor. However, these ligands are relatively unstable. Here we determined whether much more stable 2′-fluoro-modified RNA ligands can be prepared and used to study the alleviation of receptor inhibition. Two classes of 2′-fluoro-modified RNA ligands were obtained: One class binds with higher affinity to the cocaine-binding site on the closed-channel form and, as predicted by the mechanism, inhibits the receptor. The second class binds with equal or higher affinity to the cocaine-binding site on the open-channel form and, as predicted by the mechanism, does not inhibit the receptor, and does alleviate cocaine and MK-801 inhibition of the nAChR. The stability of these 2′-fluoro-RNAs expands the utility of these ligands.

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Acknowledgements

We thank Oné R. Pagan, Richard M. Hann, and Vesna A. Eterovic (Universidad Central del Caribe) for a gift of the T. californica membrane preparation, Sean Lee (Yale University School of Medicine) for doing some preliminary experiments, Susan Coombs for editorial help, and Brenda Everhart for help in preparing the manuscript. This work was supported by a grant (NS08527) awarded to G. P. H. by the National Institutes of Health, National Institute for Neurological Disease and Stroke.

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  1. Department of Molecular Biology and Genetics, Biotechnology Building, Cornell University, Ithaca, NY, 14853, USA

    Y. Cui & G. P. Hess

  2. Department of Biochemistry, Instituto de Química, Universidade de São Paulo, 05508-900 S.P., São Paulo, Brazil

    H. Ulrich

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Cui, Y., Ulrich, H. & Hess, G.P. Selection of 2′-Fluoro-modified RNA Aptamers for Alleviation of Cocaine and MK-801Inhibition of the Nicotinic Acetylcholine Receptor. J Membrane Biol 202, 137–149 (2004). https://doi.org/10.1007/s00232-004-0725-4

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