12 results on '"Dion A. Daniels"'
Search Results
2. Circadian clock component REV-ERBα controls homeostatic regulation of pulmonary inflammation
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Patricia L. Podolin, Justyna Wojno-Picon, Baoqiang Guo, Andrew S. I. Loudon, Ryan Vonslow, Ryan P. Trump, Daniel Grant, Andrew S. MacDonald, Yolanda Sanchez, Anthony William James Cooper, Marie Pariollaud, D. Heulyn Jones, Brian Bolognese, Nicholas C. O. Tomkinson, Thomas Hopwood, Stefano Bresciani, David W. Ray, William J. Zuercher, Nicola Begley, Timothy M. Willson, Sheila Brown, Toryn Poolman, James P. Tellam, Dion A. Daniels, Ben Saer, and Julie E. Gibbs
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0301 basic medicine ,Chemokine ,Pulmonology ,Neutrophils ,viruses ,Circadian clock ,SUMO protein ,Mice, Transgenic ,Inflammation ,Endogeny ,Mouse models ,Proinflammatory cytokine ,Mice ,03 medical and health sciences ,Circadian Clocks ,medicine ,Animals ,Homeostasis ,Circadian rhythm ,Innate immunity ,Innate immune system ,biology ,Sumoylation ,Pneumonia ,General Medicine ,Immunity, Innate ,Circadian Rhythm ,3. Good health ,Cell biology ,030104 developmental biology ,Nuclear Receptor Subfamily 1, Group D, Member 1 ,Proteolysis ,biology.protein ,medicine.symptom ,Research Article - Abstract
Recent studies reveal that airway epithelial cells are critical pulmonary circadian pacemaker cells, mediating rhythmic inflammatory responses. Using mouse models, we now identify the rhythmic circadian repressor REV-ERBα as essential to the mechanism coupling the pulmonary clock to innate immunity, involving both myeloid and bronchial epithelial cells in temporal gating and determining amplitude of response to inhaled endotoxin. Dual mutation of REV-ERBα and its paralog REV-ERBβ in bronchial epithelia further augmented inflammatory responses and chemokine activation, but also initiated a basal inflammatory state, revealing a critical homeostatic role for REV-ERB proteins in the suppression of the endogenous proinflammatory mechanism in unchallenged cells. However, REV-ERBα plays the dominant role, as deletion of REV-ERBβ alone had no impact on inflammatory responses. In turn, inflammatory challenges cause striking changes in stability and degradation of REV-ERBα protein, driven by SUMOylation and ubiquitination. We developed a novel selective oxazole-based inverse agonist of REV-ERB, which protects REV-ERBα protein from degradation, and used this to reveal how proinflammatory cytokines trigger rapid degradation of REV-ERBα in the elaboration of an inflammatory response. Thus, dynamic changes in stability of REV-ERBα protein couple the core clock to innate immunity.
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- 2018
3. Pharmacology of GPR55 in Yeast and Identification of GSK494581A as a Mixed-Activity Glycine Transporter Subtype 1 Inhibitor and GPR55 Agonist
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Susan Brown, Paula L. Nichols, Victoria R Terrell, Alan Wise, Simon J. Dowell, Carl Haslam, Jason L. Brown, Mumta Kassim, Penny C. Staton, Dion A. Daniels, and Andrew J. Brown
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Agonist ,Cannabinoid receptor ,medicine.drug_class ,Stereochemistry ,medicine.medical_treatment ,Saccharomyces cerevisiae ,Biology ,Pharmacology ,Piperazines ,Receptors, G-Protein-Coupled ,Glycine transporter ,Glycine Plasma Membrane Transport Proteins ,Yeasts ,medicine ,Humans ,Receptors, Cannabinoid ,Receptor ,Bone morphogenesis ,Dose-Response Relationship, Drug ,HEK 293 cells ,Ligand (biochemistry) ,HEK293 Cells ,Biochemistry ,Molecular Medicine ,Cannabinoid - Abstract
GPR55 is a G protein-coupled receptor activated by L-α-lysophosphatidylinositol and suggested to have roles in pain signaling, bone morphogenesis, and possibly in vascular endothelial cells. It has affinity for certain cannabinoids (molecules that interact with the cannabinoid CB(1) and CB(2) receptors), but investigation of its functional role in cell-based systems and in tissue has been limited by a lack of selective pharmacological tools. Here, we present our characterization of GPR55 in the yeast Saccharomyces cerevisiae and in human embryonic kidney (HEK293) cells. We describe GSK494581A (1-{2-fluoro-4-[1-(methyloxy)ethyl]phenyl}-4-{[4'-fluoro-4-(methylsulfonyl)-2-biphenylyl]carbonyl}piperazine), a selective small-molecule ligand of GPR55 identified through diversity screening. GSK494581A is one of a series of benzoylpiperazines originally identified and patented as inhibitors of the glycine transporter subtype 1 (GlyT1). The structure-activity relationship between GPR55 and GlyT1 is divergent across this series. The most GPR55-selective example is GSK575594A (3-fluoro-4-(4-{[4'-fluoro-4-(methylsulfonyl)-2-biphenylyl]carbonyl}-1-piperazinyl)aniline), which is approximately 60-fold selective for GPR55 (pEC(50) = 6.8) over GlyT1 (pIC(50) = 5.0). Several exemplars with activity at GPR55 and GlyT1 have been profiled at a broad range of other molecular targets and are inactive at cannabinoid receptors and all other targets tested. The benzoylpiperazine agonists activate human GPR55 but not rodent GPR55, suggesting that the relatively low level of sequence identity between these orthologs (75%) translates to important functional differences in the ligand-binding site.
- Published
- 2011
4. Biochemical characterization of different conformational states of the Sf9 cell-purified p53His175 mutant protein
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Dion A. Daniels, Pascale Cohen, David P. Lane, and Ted R. Hupp
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p53 ,Electrophoresis ,HMG-box ,Protein Conformation ,Biophysics ,Enzyme-Linked Immunosorbent Assay ,Transfection ,Biochemistry ,DDB1 ,His175 mutation ,Structural Biology ,Mutant protein ,Protein A/G ,Tumor Cells, Cultured ,Genetics ,Humans ,Histidine ,Electrophoretic mobility shift assay ,Conformation ,DNA binding ,Molecular Biology ,biology ,Binding protein ,Mutant ,Antibodies, Monoclonal ,Cell Biology ,Molecular biology ,DNA-Binding Proteins ,DNA binding site ,Mutagenesis ,biology.protein ,Tumor Suppressor Protein p53 ,Baculoviridae ,Protein Binding ,Binding domain - Abstract
In this study, we expressed and purified the p53 mutant encoded by the His175 allele (p53His175) in a baculovirus expression system in order to study the folding and the DNA binding activity of the protein. A two-site ELISA revealed that purified p53His175 protein preferentially displayed a PAb1620 conformation, which appeared to be not sufficient to interact specifically with DNA. The cryptic DNA binding activity of this mutant was then investigated by electrophoretic mobility shift assay in the presence of anti-p53 antibodies, and shown to be refractory to significant activation by PAb421 (a potent allosteric activator of wild-type p53’s DNA binding activity). Nevertheless, p53His175 DNA binding was regulated by antibodies targeting the N-terminal region of the protein. Furthermore, while the protein preferentially displayed a PAb1620 conformation, our data suggested the existence of an equilibrium between at least two folding states of the protein (PAb1620 and PAb240 conformations). A model rationalizing the conformation, antibody-interacting ability and DNA binding regulation potential of p53His175 is presented.
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- 1999
5. Phage Peptide Libraries
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Dion A. Daniels and David P. Lane
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chemistry.chemical_classification ,Phage display ,medicine.drug_class ,viruses ,Phagemid ,Peptide ,Biopanning ,Biology ,Monoclonal antibody ,Molecular biology ,Genome ,General Biochemistry, Genetics and Molecular Biology ,Insert (molecular biology) ,chemistry ,Biochemistry ,medicine ,Binding site ,Molecular Biology - Abstract
Filamentous phage particles have been central in the construction of libraries displaying vast numbers of random peptides. These random peptides can be antigenically presented as fusions to coat proteins III and VIII of the phage. The isolation of ligate-reactive phage from an immense background of nonspecific phage is achieved by the biopanning process. Enrichment of reactive phage relative to unreactive phage occurs with alternate rounds of affinity selection to the desired molecular target and amplification of the specifically bound phage. This allows the isolation of rare binding species contained in the phage peptide libraries. Each phage particle contains the information in its genome pertaining to the type of random peptide insert displayed. Hence, the identification of binding motifs displayed on ligate-reactive phage is revealed by sequencing the relevant insert site in the phage genome. Phage peptide libraries have been used to isolate ligands to an array of protein ligates. The libraries have proved particularly effective in defining the binding sites of monoclonal antibodies and to some extent polyclonal sera. The analysis of the peptide insert sequences of a number of different clones of antibody binding phage can reveal a great deal about the nature and restriction of the amino acid residues critical for the antibody-antigen interaction.
- Published
- 1996
6. The characterisation of p53 binding phage isolated from phage peptide display libraries
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David P. Lane and Dion A. Daniels
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Phage display ,Polymers ,Protein Conformation ,medicine.drug_class ,viruses ,Phagemid ,Molecular Sequence Data ,Restriction Mapping ,Enzyme-Linked Immunosorbent Assay ,Peptide ,Antigen-Antibody Complex ,Spodoptera ,Biology ,Monoclonal antibody ,Binding, Competitive ,Mice ,Protein structure ,Antibody Specificity ,Structural Biology ,medicine ,Animals ,Humans ,Bacteriophages ,Genomic library ,Amino Acid Sequence ,Antigens, Viral, Tumor ,Molecular Biology ,Peptide sequence ,Conserved Sequence ,chemistry.chemical_classification ,Genomic Library ,Antibodies, Monoclonal ,Molecular biology ,Recombinant Proteins ,chemistry ,Mutation ,Tumor Suppressor Protein p53 ,Peptides ,Baculoviridae ,P53 binding - Abstract
Peptide phage display libraries were screened for peptides that bind to the tumour suppressor protein, human p53. Three p53 binding peptides were isolated respectively from hexamer (6-mer), dodecamer (12-mer) and icosomer (20-mer) libraries. We have characterised their interaction with p53 in detail. The phage appear to bind regions on native p53 common between mouse and man. Two conformation-specific anti-p53 monoclonal antibodies were used to dissect the phage-p53 interaction, the phage were found to preferentially bind the PAb1620-p53 conformation rather than the PAb240-p53 conformation. Mapping experiments indicated the C-terminal 30 amino acid residues of p53 were dispensable for phage binding and that the binding of SV40 large T-antigen and the phage were not mutually exclusive. Interestingly the phage were seen to exhibit differential binding to wild-type human p53 over the two point mutant p53 proteins, His175 and Trp248. Ultimately the phage appear to selectively target native wild-type p53, mimicking the specificity of SV40 large T-antigen. The ability to target specific sub-populations of p53 could be an important step in the development of therapeutics for the treatment of p53-based human malignancy.
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- 1994
7. Antibodies that identify only the active conformation of G(i) family G protein alpha subunits
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J. Robert Lane, Chris Plumpton, Alan Wise, Graeme Milligan, Stephen Edward Rees, Ian Kinghorn, David Henderson, Dion A. Daniels, and Ben Powney
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GTPase-activating protein ,G protein ,Protein Conformation ,Protein subunit ,Mutation, Missense ,GTP-Binding Protein alpha Subunits, Gi-Go ,Biochemistry ,Guanosine Diphosphate ,Antibodies ,Mice ,Heterotrimeric G protein ,Protein A/G ,Genetics ,Animals ,Molecular Biology ,G protein-coupled receptor ,G protein-coupled receptor kinase ,Hybridomas ,biology ,Receptors, Dopamine D2 ,Molecular biology ,GTP-Binding Protein alpha Subunits ,G beta-gamma complex ,biology.protein ,Guanosine Triphosphate ,Biotechnology - Abstract
Production of antisera able to recognize individual heterotrimeric G protein alpha subunits resulted in rapid expansion of information on their distribution and function. However, no antibodies that specifically recognize the active state have been available. Four-way primary screening of 763 hybridomas generated from mice immunized with guanosine 5'-O-(3-thio)triphosphate-loaded G alpha(i1) and isolated using an automated robotic colony picker identified three antibodies that interacted with the constitutively active, Q(204)L, mutant but neither the constitutively inactive, G(203)A, mutant nor wild-type G alpha(i1). This profile extended to other closely related G(i) family G proteins but not to the less closely related G alpha(s) and G alpha(q)/G alpha(11) families. Each antibody was, however, also able to identify wild-type, GDP-bound G(i) family G proteins in the presence of fluoroaluminate, which mimics the presence of the terminal phosphate of GTP and hence generates an active/transition state conformation. Stimulation of cells coexpressing a wild-type G alpha(i) subunit and the dopamine D2 receptor with the agonist ligand nor-apomorphine also allowed these conformationally selective antibodies to bind the G protein. Such reagents allow the specific identification of activated G proteins in a native environment and may allow the development of label-free screening assays for G protein-coupled receptor-mediated activation of G(i) family G proteins.
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- 2008
8. In vitro selection of RNA aptamers that block CCL1 chemokine function
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Dion A. Daniels, Trevor D. Chapman, Katy L. Gearing, Martin L Marro, and David P. Andrew
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Aptamer ,Molecular Sequence Data ,Biophysics ,CCL1 ,Biology ,CCR8 ,In Vitro Techniques ,Ligands ,Biochemistry ,Chemokine CCL1 ,Mice ,In vivo ,Animals ,Humans ,Molecular Biology ,Inflammation ,Base Sequence ,Chemotactic Factors ,Dose-Response Relationship, Drug ,RNA ,Chemotaxis ,Cell Biology ,Ligand (biochemistry) ,Molecular biology ,Cell biology ,Chemokines, CC ,Nucleic Acid Conformation ,Chemokines ,Systematic evolution of ligands by exponential enrichment - Abstract
CCL1, the CCR8 ligand, is a CC chemokine secreted by activated monocytes and lymphocytes and is a potent chemoattractant for these cell types. The in vivo role of the CCL1/CCR8 axis in Th2-mediated inflammation is far from clear. Ligand neutralisation studies reported discrepancies in the effect of CCL1/CCR8 and CCR8 knockout studies showed very different insights into the functional role of the CCR8. To further study the biological function of CCL1, we focused on the generation and characterisation of RNA aptamers. We report here the in vitro isolation of the first nuclease resistant and selective RNA aptamer (T48) with high-binding affinity for human and mouse CCL1. The T48 aptamer but not a random control aptamer antagonises CCL1 function in a dose-dependent fashion in both heparin binding and chemotaxis assays. To our knowledge, the T48 aptamer constitutes one of the most potent CCL1 antagonists reported to date and is an excellent tool to dissect CCL1-specific function in vivo. The T48 aptamer may also have potential as new generation of therapeutic tools.
- Published
- 2006
9. Identification of potent and selective RNA antagonists of the IFN-gamma-inducible CXCL10 chemokine
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Dion A. Daniels, Trevor D. Chapman, Kalpana Patel, Zining Wu, Janet L. Smith, Anne McNamee, Katy L. Gearing, David P. Andrew, Ming S Jiang, and Martin L Marro
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Chemokine ,Receptors, CXCR3 ,Aptamer ,Molecular Sequence Data ,CHO Cells ,Biology ,CXCR3 ,Ligands ,Biochemistry ,Polyethylene Glycols ,Interferon-gamma ,Mice ,Cell Line, Tumor ,Cricetinae ,CXCL10 ,Animals ,Humans ,Binding Sites ,Base Sequence ,Chinese hamster ovary cell ,RNA ,RNA-Binding Proteins ,hemic and immune systems ,respiratory system ,Molecular biology ,In vitro ,Cell biology ,Rats ,Chemokine CXCL10 ,Cell Migration Inhibition ,PEGylation ,biology.protein ,Receptors, Chemokine ,Chemokines, CXC ,Signal Transduction - Abstract
CXCL10 (also known as IP-10 in humans and CRG-2 in mice) is a nonglycosylated chemokine and a member of the non-ELR CXC chemokine subfamily implicated in a variety of inflammatory conditions. The role of CXCL10 in different disease states still requires clarification, and new approaches are necessary to better understand its biological function. We report here the isolation of a series of nuclease-resistant RNA aptamers that act to antagonize human CXCL10 function in a number of in vitro and cell-based assays. The two most potent aptamers identified were highly selective for human CXCL10. A further aptamer was identified that antagonized both the human and the mouse CXCL10. A combination of a molecular-biology-based truncation and solid-phase synthesis enabled the truncation of one of the aptamers from 71 to 34 nucleotides. This was followed by PEGylation, 3' capping, and further stabilization of the RNA aptamer, while its high potency was maintained. These aptamers could be utilized as powerful target validation tools and may also have therapeutic potential. To our knowledge, the CXCL10 aptamers generated are the most potent antagonists of CXCL10/CXCR3 signaling reported to date.
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- 2005
10. A tenascin-C aptamer identified by tumor cell SELEX: Systematic evolution of ligands by exponential enrichment
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Larry Gold, Brian Hicke, Dion A. Daniels, Kristine M. Swiderek, and Hang Chen
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Aptamer ,Molecular Sequence Data ,DNA, Single-Stranded ,Enzyme-Linked Immunosorbent Assay ,Computational biology ,Biology ,medicine.disease_cause ,Ligands ,Mass Spectrometry ,chemistry.chemical_compound ,medicine ,Tumor Cells, Cultured ,Humans ,Amino Acid Sequence ,Genetics ,Multidisciplinary ,Base Sequence ,Oligonucleotide ,Tenascin C ,Membrane Proteins ,Tenascin ,Templates, Genetic ,Biological Sciences ,Biological Evolution ,Peptide Fragments ,chemistry ,Membrane protein ,Oligodeoxyribonucleotides ,SELEX Aptamer Technique ,biology.protein ,Carcinogenesis ,Glioblastoma ,DNA ,Systematic evolution of ligands by exponential enrichment ,Chromatography, Liquid - Abstract
The targeting of molecular repertoires to complex systems rather than biochemically pure entities is an accessible approach that can identify proteins of biological interest. We have probed antigens presented by a monolayer of tumor cells for their ability to interact with a pool of aptamers. A glioblastoma-derived cell line, U251, was used as the target for systematic evolution of ligands by exponential enrichment by using a single-stranded DNA library. We isolated specifically interacting oligonucleotides, and biochemical strategies were used to identify the protein target for one of the aptamers. Here we characterize the interaction of the DNA aptamer, GBI-10, with tenascin-C, an extracellular protein found in the tumor matrix. Tenascin-C is believed to be involved in both embryogenesis and oncogenesis pathways. Systematic evolution of ligands by exponential enrichment appears to be a successful strategy for the a priori identification of targets of biological interest within complex systems.
- Published
- 2003
11. The Orphan G protein-coupled receptors GPR41 and GPR43 are activated by propionate and other short chain carboxylic acids
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Shelagh Wilson, Fiona H. Marshall, Alan Wise, Jay C. Strum, Andrew J. Brown, Ian Kinghorn, Mark J. Wigglesworth, Paul R. Murdock, Diane M. Ignar, Susan M. Goldsworthy, Lili Tcheang, Julie C. Holder, Nicholas B. Pike, Neil J. Fraser, Michelle M. Eilert, Ashley A. Barnes, Philip G. Szekeres, Dion A. Daniels, Klaudia Steplewski, Alison I. Muir, Simon J. Dowell, and Steve M. Foord
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Agonist ,medicine.drug_class ,Molecular Sequence Data ,Carboxylic Acids ,Guanosine ,Receptors, Cell Surface ,Butyrate ,Biology ,Biochemistry ,Receptors, G-Protein-Coupled ,chemistry.chemical_compound ,Xenopus laevis ,medicine ,Animals ,Humans ,G protein-coupled inwardly-rectifying potassium channel ,Amino Acid Sequence ,Receptor ,Molecular Biology ,G protein-coupled receptor ,DNA Primers ,chemistry.chemical_classification ,GPR120 ,Cell Biology ,Molecular biology ,Immunohistochemistry ,Recombinant Proteins ,chemistry ,Propionate ,Propionates - Abstract
GPR41 and GPR43 are related members of a homologous family of orphan G protein-coupled receptors that are tandemly encoded at a single chromosomal locus in both humans and mice. We identified the acetate anion as an agonist of human GPR43 during routine ligand bank screening in yeast. This activity was confirmed after transient transfection of GPR43 into mammalian cells using Ca(2+) mobilization and [(35)S]guanosine 5'-O-(3-thiotriphosphate) binding assays and by coexpression with GIRK G protein-regulated potassium channels in Xenopus laevis oocytes. Other short chain carboxylic acid anions such as formate, propionate, butyrate, and pentanoate also had agonist activity. GPR41 is related to GPR43 (52% similarity; 43% identity) and was activated by similar ligands but with differing specificity for carbon chain length, with pentanoate being the most potent agonist. A third family member, GPR42, is most likely a recent gene duplication of GPR41 and may be a pseudogene. GPR41 was expressed primarily in adipose tissue, whereas the highest levels of GPR43 were found in immune cells. The identity of the cognate physiological ligands for these receptors is not clear, although propionate is known to occur in vivo at high concentrations under certain pathophysiological conditions.
- Published
- 2002
12. Generation of RNA aptamers to the G-protein-coupled receptor for neurotensin, NTS-1
- Author
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Awinder K. Sohal, Stephen Rees, Reinhard Grisshammer, and Dion A. Daniels
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Sequence Analysis, RNA ,Aptamer ,Recombinant Fusion Proteins ,Biophysics ,Temperature ,RNA ,Cell Biology ,Biology ,Biochemistry ,Rats ,chemistry.chemical_compound ,chemistry ,GTP-Binding Proteins ,Animals ,Receptors, Neurotensin ,Neurotensin receptor ,Signal transduction ,Receptor ,Molecular Biology ,Integral membrane protein ,Neurotensin ,G protein-coupled receptor - Abstract
G-protein-coupled receptors (GPCRs) are integral membrane proteins involved in signal transduction and constitute major drug targets for disease therapy. Aptamers, which are globular RNA or DNA molecules evolved to specifically bind a target, could represent a valuable tool with which to probe the role of such receptors in normal tissue and disease pathology and for cocrystallization with receptors for structure determination by X-ray crystallography. Using the bacterially expressed rat neurotensin receptor NTS-1 as an example, we describe a strategy for the generation of GPCR-specific RNA aptamers. Seven rounds of a "subtractive," paramagnetic bead-based selection protocol were used to enrich for neurotensin receptor-specific aptamers, while circumventing the evolution of aptamers reactive to minor protein contaminants. Representatives of each aptamer family were analyzed in Escherichia coli membrane nitrocellulose filter binding assays. Eight aptamers demonstrated specificity for the neurotensin receptor. One aptamer, P19, was characterized in detail and shown to bind to both the rat receptor and the human receptor with nanomolar affinity. P19 was also shown to interact with rat neurotensin receptor expressed in CHO cells, in both membrane preparations and intact cells. P19 represents the first example of a GPCR-specific RNA aptamer.
- Published
- 2002
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