Anti-alprenolol anti-idiotypic antibodies bind to beta-adrenergic receptors and modulate catecholamine-sensitive adenylate cyclase.

Abstract

To better understand the interaction between f3-adrenergic ligands and their specific membrane receptors, we have undertaken to isolate and purify the various components intervening in the catecholamine-mediated activation of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1]. For this purpose we have developed an affinity chromatography method that allows physical separation of the functional cyclase and f3-adrenergic receptors from turkey erythrocyte membranes (1).Various biochemical and pharmacological properties of the receptors were described (2). The minute amounts of protein prepared by this method and the risk of losing essential components of the receptor cyclase complex during the membrane purification and solubilization steps prompted us to develop a second line of investigation based on immunological methods.Anti-receptor antibodies can be obtained by immunization of rabbits and mice with purified ,B-adrenergic receptor, but most of the antibodies react with parts of the molecule other than the binding site (ref. 3; unpublished data). This is in agreement with observations made in other systems: animals do not make anti-receptor binding site antibodies upon immunization with insulin, acetylcholine, or thyrotropin receptors despite the fact that in pathological conditions such as autoimmune diabetes (4), myasthenia gravis (5), Graves disease (6), and 02-adrenergic hyporesponsiveness in allergic rhinitis (7) anti-binding site antibodies probably constitute an important basis for the etiopathogeny.

An antiidiotypic antibody that recognizes the beta-adrenergic receptor.

“…Our antiidiotype behaved in a manner most consistent with competitive inhibition, determined both by saturation binding analysis and as assessed by isoproterenol-mediated adenylate cyclase activation. This contrasts with the results of Schreiber et al (4). By Scatchard analysis, their antibody displayed the characteristics of noncompetitive antagonism.…”

“…lB 14 16 18 20 BOUND (fmo/) FIGURE 1 Effect of antiidiotypic antiserum on (3-receptor function as assessed by Scatchard analysis. Turkey erythrocyte membranes were incubated with increasing concentrations of (-)-[3Hlalprenolol, [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] Fig. 2.…”

“…Such an approach has already been successfully applied with insulin, a far more complex ligand than alprenolol, in the identification of insulin receptors on cell membranes (3). More recently, Schreiber et al (4) have also reported on the development of antiidiotypic antibodies that recognize the ,-receptor. Important contrasts between our results are evident.…”

Anti-dihydropyridine Antibodies Exhibit [3H]Nitrendipine Binding Properties Similar to the Membrane Receptor for the 1,4-Dihydropyridine Ca2+ Channel Antagonists

“…Anti-idiotypic antibodies have been very useful in studying receptor structure and function, as these antibodies bind to the ligand binding site on the receptor and in some cases can modify receptor function (21). Anti-ligand antibodies (20,21,23,(26)(27)(28)(29) have been used to study structure-activity relationships between the ligand and antibody, and as specific molecules that can compete with the receptor for the ligand.…”

“…Considerable advances have been made in recent years in the structural and functional characterization of membrane receptors and ion channels using immunelogical probes (20)(21)(22)(23)(24)(25)(26), Antibodies have been raised against membrane receptors or ion channels directly by immunization with purified membrane proteins, or indirectly by immunization with anti-ligand antibodies to produce anti-idiotypic antibodies. Anti-idiotypic antibodies have been very useful in studying receptor structure and function, as these antibodies bind to the ligand binding site on the receptor and in some cases can modify receptor function (21).…”