Use of electrofocusing for the analysis and purification of turkey erythrocytes beta 1-adrenoceptors.
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.
Marbach
I
I
Department of Biological Chemistry, Hebrew University of Jerusalem, Israel.
Levitzki
A
A
D000180:Acrylic Resins; D000818:Animals; D001667:Binding, Competitive; D002846:Chromatography, Affinity; D004072:Digitonin; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007457:Iodine Radioisotopes; D007525:Isoelectric Focusing; D010869:Pindolol / Q000031:analogs & derivatives / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000302:isolation & purification* / Q000378:metabolism; D014422:Turkeys / Q000097:blood
D000180:Acrylic Resins; D007457:Iodine Radioisotopes; D011943:Receptors, Adrenergic, beta; C016680:polyacrylamide gels; C047975:cyanopindolol; D010869:Pindolol; D004072:Digitonin