Abstract
Antialprenolol rabbit antibodies were fractionated on an acebutolol affinity resin, followed by L-propranolol elution so as to separate a class of binding sites that mimic the beta-adrenergic receptor. Allotype-identicaL rabbits were immunized with this fraction. After 6 mo, antisera exhibited antiidiotypic activity inhibiting [3H]alprenolol binding to the original antibody and to rabbit antiacebutolol antibodies, which had a spectrum of ligand-binding properties identical to the original idiotype. Those antisera demonstrating the original idiotype. Those antisera demonstrating the most potent antiidiotypic activity also blocked [3H]alprenolol binding to the beta-adrenergic receptor of turkey membrane, canine pulmonary membrane, and rat reticulocyte. An idiotype affinity-purified fraction showed similar activity, inhibiting beta-receptor binding with a calculated dissociation constant (KD) of 53 nM. Isoproterenol-mediated adenylate cyclase activity was also inhibited in a competitive manner. The universality of recognition of these antiidiotypic antisera indicate that the three-dimensional structure of a receptor's binding site can be modeled by a subset of an elicited antibody population.
MeSH Terms
Acebutolol/immunology
Alprenolol/immunology
Animals
Antibody Affinity
Antibody Formation
Binding Sites, Antibody
Immunization
Immunoglobulin Idiotypes
Propranolol/pharmacology
Rabbits
Receptors, Adrenergic/immunology
Receptors, Adrenergic, beta/immunology
Chemicals
Immunoglobulin Idiotypes
Receptors, Adrenergic
Receptors, Adrenergic, beta
Acebutolol
Alprenolol
Propranolol
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Homcy C J
Rockson S G
Haber E
References (18)
18 references, click to expand
-
Plasma membranes of the rat liver. Isolation and enzymatic characterization of a fraction rich in bile canaliculi.
J Cell Biol. 1969 Apr;41(1):124-32
PMID: 4304740
-
An enzymic method for the trace iodination of immunoglobulins and other proteins.
Biochem J. 1969 Jun;113(2):299-305
PMID: 4185494
-
A modified method for the isolation of the plasma membrane from rat liver.
Biochim Biophys Acta. 1970 Jan 6;196(1):1-9
PMID: 4312696
-
Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads.
J Biol Chem. 1970 Jun;245(12):3059-65
PMID: 5432796
-
Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction.
Biochem Pharmacol. 1973 Dec 1;22(23):3099-108
PMID: 4202581
-
A highly sensitive adenylate cyclase assay.
Anal Biochem. 1974 Apr;58(2):541-8
PMID: 4827395
-
Demonstration of the hydrophilic character of adenylate cyclase following hydrophobic resolution on immobilized alkyl residues. Critical role of alkyl chain length.
J Biol Chem. 1977 Dec 25;252(24):8957-64
PMID: 925032
-
Use of anti-idiotypic antibodies as cell-surface receptor probes.
Proc Natl Acad Sci U S A. 1978 May;75(5):2443-7
PMID: 307766
-
The production and characterization of antibodies against beta-adrenergic antagonists.
Biochem Pharmacol. 1978;27(11):1527-32
PMID: 29641
-
The role of the guanine nucleotide exchange reaction in the regulation of the beta-adrenergic receptor and in the actions of catecholamines and cholera toxin on adenylate cyclase in turkey erythrocyte membranes.
J Biol Chem. 1980 Feb 10;255(3):988-95
PMID: 6243304
-
Hybridoma proteins expressing the predominant idiotype of the antiazophenylarsonate response of A/J mice.
Proc Natl Acad Sci U S A. 1980 Feb;77(2):1120-4
PMID: 6767242
-
Beta receptor occupancy. Assessment in the intact animal.
J Clin Invest. 1980 May;65(5):1111-8
PMID: 6102571
-
Anti-alprenolol antibodies in the rabbit. A new probe for the study of beta-adrenergic receptor interaction.
Circ Res. 1980 Jun;46(6):808-13
PMID: 6247081
-
Photoaffinity label for the beta-adrenergic receptor: synthesis and effects on isoproterenol-stimulated adenylate cyclase.
Proc Natl Acad Sci U S A. 1980 Aug;77(8):4449-53
PMID: 6254025
-
Cellular mechanisms of impaired adrenergic responsiveness in neonatal dogs.
J Clin Invest. 1981 Feb;67(2):319-27
PMID: 6257759
-
Anti-alprenolol anti-idiotypic antibodies bind to beta-adrenergic receptors and modulate catecholamine-sensitive adenylate cyclase.
Proc Natl Acad Sci U S A. 1980 Dec;77(12):7385-9
PMID: 6261254
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
The hydrolysis of rabbit y-globulin and antibodies with crystalline papain.
Biochem J. 1959 Sep;73:119-26
PMID: 14434282