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Use of electrofocusing for the analysis and purification of turkey erythrocytes β 1 ‐adrenoceptors
Author(s) -
Marbach Irit,
Levitzki Alexander
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)81381-4
Subject(s) - isoelectric focusing , polyacrylamide , chromatography , electroelution , polyacrylamide gel electrophoresis , chemistry , receptor , yield (engineering) , digitonin , biochemistry , membrane , enzyme , materials science , polymer chemistry , metallurgy
We show that following one cycle of alprenolol affinity chromatography of turkey erythrocyte β 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 β‐adrenoceptor is 75 ± 3 %. In addition, we are able to demonstrate that receptor‐binding assays can be performed directly on the polyacrylamide gel, using 125 I‐cyanopindolol. This method can be employed for minute quantities of receptor which is an advantage when one wishes to characterize rapidly the β‐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 β 1 ,‐adrenoceptor on immobiline polyacrylamide gels and the ability to purify only partially the receptor on these gels.