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Modulation of the M2 muscarinic acetylcholine receptor activity with monoclonal anti-M2 receptor antibody fragments

Item Type:Article
Title:Modulation of the M2 muscarinic acetylcholine receptor activity with monoclonal anti-M2 receptor antibody fragments
Creators Name:Peter, J.C. and Wallukat, G. and Tugler, J. and Maurice, D. and Roegel, J.C. and Briand, J.P. and Hoebeke, J.
Abstract:Antibodies directed against the second extracellular loop of G protein-coupled receptors are known to have functional activities. From a partial agonist monoclonal antibody directed against the M2 muscarinic receptor, we constructed and produced a single chain variable fragment with high affinity for its target epitope. The fragment is able to recognize its receptor on Chinese hamster ovary cells transfected with the M2 muscarinic acetylcholine receptor to block the effect of carbachol on this receptor and to exert an inverse agonist activity on the basal activity of the receptor. The antibody fragment is also able to increase the basal rhythm of cultured neonatal rat cardiomyocytes and to inhibit in a non-competitive manner the negative chronotropic effect of carbachol. This antibody fragment is able to exert its inverse agonist activity in vivo on mouse heart activity. The immunological strategy presented here could be useful to develop specific allosteric inverse agonist reagents for G protein-coupled receptors.
Keywords:Allosteric Site, Amino Acid Sequence, Amino Acid Sequence Homology, Base Sequence, Carbachol, Cardiac Myocytes, CHO Cells, Cholinergic Receptors, Competitive Binding, Cricetinae, Cultured Cells, Drug Dose-Response Relationship, Epitopes, Escherichia coli, Immunoglobulin Fab Fragments, Immunohistochemistry, Immunologic Dose-Response Relationship, Molecular Models, Molecular Sequence Data, Monoclonal Antibodies, Muscarinic M2 Receptor, Nucleotides, Peptides, Polyacrylamide Gel Electrophoresis, Surface Plasmon Resonance, Tertiary Protein Structure, Time Factors, Transfection, Western Blotting, Animals, Mice, Rats
Source:Journal of Biological Chemistry
ISSN:0021-9258
Publisher:American Society for Biochemistry and Molecular Biology (U.S.A.)
Volume:279
Number:53
Page Range:55697-55706
Date:31 December 2004
Official Publication:https://doi.org/10.1074/jbc.M407213200
PubMed:View item in PubMed

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