digitonin and cyanopindolol

We show that following one cycle of alprenolol affinity chromatography of turkey erythrocyte beta 1-adrenoceptors, electrofocusing on polyacrylamide gels in digitonin, followed by electroelution, results in complete receptor purification. The overall yield from the electrofocusing-electroelution step of turkey erythrocyte beta-adrenoceptor is 75 +/- 3%. In addition, we are able to demonstrate that receptor-binding assays can be performed directly on the polyacrylamide gel, using 125I-cyanopindolol. This method can be employed for minute quantities of receptor which is an advantage when one wishes to characterize rapidly the beta-adrenoceptor in its native state from tissues that may be available only in limited amounts. We also report, for comparison, on the behavior of the turkey erythrocyte beta 1-adrenoceptor on immobiline polyacrylamide gels and the ability to purify only partially the receptor on these gels.

Topics: Acrylic Resins; Animals; Binding, Competitive; Chromatography, Affinity; Digitonin; Electrophoresis, Polyacrylamide Gel; Erythrocytes; Hydrogen-Ion Concentration; Iodine Radioisotopes; Isoelectric Focusing; Pindolol; Receptors, Adrenergic, beta; Turkeys