Mammalian beta-adrenoceptors: concomitant biospecific elution with protein kinase activity from Sepharose-alprenolol.
The mammalian beta-adrenoceptors were solubilized from crude guinea pig lung membranes with the plant glycoside digitonin. The solubilized receptors were absorbed by affinity gels with (+/-)alprenolol as ligand and could be eluted biospecifically in good yield with either agonists or antagonists. A more than 100-fold purification can be achieved in a one-step procedure. Antagonist-eluted beta-adrenoceptors have affinities for antagonists similar to those of particulate or solubilized receptors. Their binding constants for agonists, however, are comparable to those of the particulate receptor. A cAMP-independent protein kinase activity was eluting concomitantly with the beta-adrenoceptors. The protein kinase phosphorylated endogenous substrates with 50 000 and greater than 150 000 subunit molecular weight.
D000262:Adenylyl Cyclases / Q000378:metabolism; D000526:Alprenolol; D000818:Animals; D002846:Chromatography, Affinity; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008168:Lung / Q000378:metabolism; D008566:Membranes / Q000187:drug effects; D011485:Protein Binding; D011494:Protein Kinases / Q000302:isolation & purification*; D011941:Receptors, Adrenergic / Q000302:isolation & purification*; D011943:Receptors, Adrenergic, beta / Q000302:isolation & purification*
D011941:Receptors, Adrenergic; D011943:Receptors, Adrenergic, beta; D000526:Alprenolol; D011494:Protein Kinases; D000262:Adenylyl Cyclases