%0 Journal Article %1 Mayer2008 %A Mayer, G. %A Wulffen, B. %A Huber, C. %A Brockmann, J. %A Flicke, B. %A Neumann, L. %A Hafenbradl, D. %A Klebl, B. M. %A Lohse, M. J. %A Krasel, C. %A Blind, M. %D 2008 %J RNA %K & 2/*antagonists Acid Aptamer Aptamers, Base Conformation DNA Data Enzyme G-Protein-Coupled Humans Inhibitors/*pharmacology Kinase Molecular Nucleic Nucleotide/chemistry/genetics/pharmacology Primers/genetics Protein Proteins/antagonists RNA/chemistry/genetics/*pharmacology Receptor Recombinant SELEX Sequence Structure, Technique Tertiary inhibitors/chemistry/genetics/metabolism %N 3 %P 524-34 %T An RNA molecule that specifically inhibits G-protein-coupled receptor kinase 2 in vitro %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18230760 %V 14 %X G-protein-coupled receptors are desensitized by a two-step process. In a first step, G-protein-coupled receptor kinases (GRKs) phosphorylate agonist-activated receptors that subsequently bind to a second class of proteins, the arrestins. GRKs can be classified into three subfamilies, which have been implicated in various diseases. The physiological role(s) of GRKs have been difficult to study as selective inhibitors are not available. We have used SELEX (systematic evolution of ligands by exponential enrichment) to develop RNA aptamers that potently and selectively inhibit GRK2. This process has yielded an aptamer, C13, which bound to GRK2 with a high affinity and inhibited GRK2-catalyzed rhodopsin phosphorylation with an IC50 of 4.1 nM. Phosphorylation of rhodopsin catalyzed by GRK5 was also inhibited, albeit with 20-fold lower potency (IC50 of 79 nM). Furthermore, C13 reveals significant specificity, since almost no inhibitory activity was detectable testing it against a panel of 14 other kinases. The aptamer is two orders of magnitude more potent than the best GRK2 inhibitors described previously and shows high selectivity for the GRK family of protein kinases.

@article{Mayer2008, abstract = {G-protein-coupled receptors are desensitized by a two-step process. In a first step, G-protein-coupled receptor kinases (GRKs) phosphorylate agonist-activated receptors that subsequently bind to a second class of proteins, the arrestins. GRKs can be classified into three subfamilies, which have been implicated in various diseases. The physiological role(s) of GRKs have been difficult to study as selective inhibitors are not available. We have used SELEX (systematic evolution of ligands by exponential enrichment) to develop RNA aptamers that potently and selectively inhibit GRK2. This process has yielded an aptamer, C13, which bound to GRK2 with a high affinity and inhibited GRK2-catalyzed rhodopsin phosphorylation with an IC50 of 4.1 nM. Phosphorylation of rhodopsin catalyzed by GRK5 was also inhibited, albeit with 20-fold lower potency (IC50 of 79 nM). Furthermore, C13 reveals significant specificity, since almost no inhibitory activity was detectable testing it against a panel of 14 other kinases. The aptamer is two orders of magnitude more potent than the best GRK2 inhibitors described previously and shows high selectivity for the GRK family of protein kinases.}, added-at = {2010-12-14T18:12:02.000+0100}, author = {Mayer, G. and Wulffen, B. and Huber, C. and Brockmann, J. and Flicke, B. and Neumann, L. and Hafenbradl, D. and Klebl, B. M. and Lohse, M. J. and Krasel, C. and Blind, M.}, biburl = {https://www.bibsonomy.org/bibtex/25ca3beff4646746513391f9316c62496/pharmawuerz}, endnotereftype = {Journal Article}, interhash = {26aa766ecace035729b581071a9665c0}, intrahash = {5ca3beff4646746513391f9316c62496}, issn = {1469-9001 (Electronic) 1469-9001 (Linking)}, journal = {RNA}, keywords = {& 2/*antagonists Acid Aptamer Aptamers, Base Conformation DNA Data Enzyme G-Protein-Coupled Humans Inhibitors/*pharmacology Kinase Molecular Nucleic Nucleotide/chemistry/genetics/pharmacology Primers/genetics Protein Proteins/antagonists RNA/chemistry/genetics/*pharmacology Receptor Recombinant SELEX Sequence Structure, Technique Tertiary inhibitors/chemistry/genetics/metabolism}, month = Mar, note = {Mayer, Gunter Wulffen, Bernhard Huber, Christian Brockmann, Jorg Flicke, Birgit Neumann, Lars Hafenbradl, Doris Klebl, Bert M Lohse, Martin J Krasel, Cornelius Blind, Michael In Vitro Research Support, Non-U.S. Gov't United States RNA (New York, N.Y.) RNA. 2008 Mar;14(3):524-34. Epub 2008 Jan 29.}, number = 3, pages = {524-34}, shorttitle = {An RNA molecule that specifically inhibits G-protein-coupled receptor kinase 2 in vitro}, timestamp = {2010-12-14T18:18:57.000+0100}, title = {An RNA molecule that specifically inhibits G-protein-coupled receptor kinase 2 in vitro}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18230760}, volume = 14, year = 2008 }