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Immunochemical and ...
Immunochemical and functional characterization of an agonist-like monoclonal antibody against the M2 acetylcholine receptor.
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- Elies, Rozenn (author)
- Gothenburg University,Göteborgs universitet,Wallenberglaboratoriet,Wallenberg Laboratory
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- Fu, Michael, 1963 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för invärtesmedicin,Wallenberglaboratoriet,Institute of Internal Medicine,Wallenberg Laboratory
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- Eftekhari, Pierre (author)
- Gothenburg University,Göteborgs universitet,Wallenberglaboratoriet,Wallenberg Laboratory
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- Wallukat, G (author)
- Gothenburg University,Göteborgs universitet,Wallenberglaboratoriet,Wallenberg Laboratory
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- Schulze, W (author)
- Gothenburg University,Göteborgs universitet,Wallenberglaboratoriet,Wallenberg Laboratory
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Granier, C (author)
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- Hjalmarson, Åke, 1937 (author)
- Gothenburg University,Göteborgs universitet,Wallenberglaboratoriet,Hjärt-kärlinstitutionen,Wallenberg Laboratory,Cardiovascular Institute
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- Hoebeke, Johan (author)
- Gothenburg University,Göteborgs universitet,Wallenberglaboratoriet,Wallenberg Laboratory
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(creator_code:org_t)
- 1998
- 1998
- English.
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In: European journal of biochemistry / FEBS. - 0014-2956. ; 251:3, s. 659-66
- Related links:
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https://gup.ub.gu.se...
Abstract
Subject headings
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- Monoclonal antibodies were raised against a peptide corresponding to the second extracellular loop of the M2 acetylcholine receptor. One of the monoclonal antibodies, B8E5, was selected for further characterization on the basis of its high yield, its isotype (IgG2a), its dissociation kinetics and its agonist-like activity. The epitope recognized by B8E5 corresponded to the N-terminal part of the second extracellular loop of the receptor (V-R-T-V-E-) as determined by competition immunoassays and epitope scanning. The KA of B8E5 for the target peptide was assessed by surface plasmon resonance (SPR) to be 6.5x10(7) M(-1) by equilibrium and 3.7x10(7) M(-1) by kinetic analysis. B8E5 recognized the M2 acetylcholine receptor on rat cardiac tissue. It only recognized the non-reduced receptor in immunoblots. The antibody had no effect on antagonist binding but decreased the affinity for the agonist carbachol. B8E5 decreased the beating frequency of neonatal rat cardiomyocytes. The effect was specific since it was blocked by the target peptide and the antagonist atropine. The EC50 of the antibody corresponded to the KA measured by surface plasmon resonance. The physiological effect of the antibody did not lead to desensitization. The Fab fragments had no physiological effect; subsequent addition of anti-mouse IgG however restored the physiological effect. These results confirm that the N-terminus of the second extracellular loop is a functional target for antibodies against the M2 acetylcholine receptor. They suggest that the functional epitope is only accessible in the non-reduced receptor. The antibodies act through a functional dimerization of the receptor.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Fysiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Physiology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Mikrobiologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Microbiology in the medical area (hsv//eng)
Keyword
- Amino Acid Sequence
- Animals
- Animals
- Newborn
- Antibodies
- Monoclonal
- isolation & purification
- pharmacology
- Atropine
- pharmacology
- Epitopes
- analysis
- metabolism
- Heart Ventricles
- Humans
- Immunoblotting
- Immunohistochemistry
- Kinetics
- Molecular Sequence Data
- Myocardial Contraction
- drug effects
- physiology
- Myocardium
- cytology
- metabolism
- Peptide Fragments
- chemistry
- immunology
- metabolism
- Rats
- Receptor
- Muscarinic M2
- Receptors
- Muscarinic
- analysis
- immunology
- metabolism
Publication and Content Type
- ref (subject category)
- art (subject category)
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Elies, Rozenn
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Fu, Michael, 196 ...
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Eftekhari, Pierr ...
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Wallukat, G
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Schulze, W
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Granier, C
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Hjalmarson, Åke, ...
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Hoebeke, Johan
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MEDICAL AND HEAL ...
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MEDICAL AND HEAL ...
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University of Gothenburg