Marbach I, Levitzki A
Department of Biological Chemistry, Hebrew University of Jerusalem, Israel.
FEBS Lett. 1988 Jun 6;233(1):186-90. doi: 10.1016/0014-5793(88)81381-4.
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.
我们发现，对火鸡红细胞β1 -肾上腺素能受体进行一轮阿普洛尔亲和层析，随后在洋地黄皂苷存在下于聚丙烯酰胺凝胶上进行电聚焦，再经电洗脱，可实现受体的完全纯化。火鸡红细胞β -肾上腺素能受体电聚焦 - 电洗脱步骤的总产率为75±3%。此外，我们能够证明可使用125I -氰吲哚洛尔直接在聚丙烯酰胺凝胶上进行受体结合测定。该方法可用于极少量的受体，当希望从数量有限的组织中快速鉴定天然状态的β -肾上腺素能受体时，这是一个优势。为作比较，我们还报告了火鸡红细胞β1 -肾上腺素能受体在固定化聚丙烯酰胺凝胶上的行为，以及在这些凝胶上仅能部分纯化该受体的情况。
