Use of electrofocusing for the analysis and purification of turkey erythrocytes β1‐adrenoceptors

FEBS Letters

Levitzki, Alexander¹  Marbach, Irit¹ 

[1]Department of Biological Chemistry, Institute of Life Sciences, The Hebrew University of Jerusalem, 91904 Jerusalem, Israel

关键词: β-Adrenoceptor;    Electrofocusing;    Ampholine;    Immobiline;    CYP;    cyanopindolol;    PEI;    polyethylenimine;    TEMED;    N;    N;    N′;    N′-tetramethylethylenediamine;    PMSF;    p-methylphenylsulfonyl fluoride;    DHA;    dihydroalprenolol;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(88)81381-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We show that following one cycle of alprenolol affinity chromatography of turkey erythrocyte β₁,-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 ¹²⁵I-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 β₁,-adrenoceptor on immobiline polyacrylamide gels and the ability to purify only partially the receptor on these gels.

【 授权许可】

Unknown