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某些离子型和非离子型去污剂对红细胞EDTA膜蛋白的比较提取

Comparative extraction of erythrocyte EDTA-membrane proteins by some ionic and non-ionic detergents.

作者信息

Ballestrin G, Covaz L, Scutari G

出版信息

Boll Soc Ital Biol Sper. 1980 Jun 15;56(11):1103-8.

PMID:6449956
Abstract

In order to examine whether it would be possible to obtain, by a simple extraction procedure from EDTA-erythrocyte-membranes, a partially purified preparation of the "band 3 zone" proteins, we have tested four solubilizing agents of common use. Detergents, both ionic (DOC and SDS) and non ionic (Tween 80 and Triton X-100), were not able, in our experimental conditions, to completely solubilize erythrocyte fragmented membranes which had previously been washed in EDTA-buffers. However, they were able to solubilize some of the membrane proteins, which could then be separated by SDS-PGE. The PGE densitometric profiles reported in this communication indicate that the protein mixture extracted by the ionic detergents DOC and SDS qualitatively reflects the protein composition of the membranes. Among the non ionic detergents, on the other hand, Triton X-100 appeared to be able to extract mainly one band (most probably the band 3 zone), while Tween 80 did not apparently extract any of the membrane proteins. Detergent concentrations, medium composition and experimental procedures are described in detail.

摘要

为了研究是否有可能通过从EDTA红细胞膜进行简单提取程序获得“带3区”蛋白的部分纯化制剂,我们测试了四种常用的增溶剂。在我们的实验条件下,离子型洗涤剂(脱氧胆酸钠和十二烷基硫酸钠)和非离子型洗涤剂(吐温80和曲拉通X-100)都不能完全溶解先前在EDTA缓冲液中洗涤过的红细胞破碎膜。然而,它们能够溶解一些膜蛋白,然后可以通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳将其分离。本通讯中报道的聚丙烯酰胺凝胶电泳光密度分析图谱表明,离子型洗涤剂脱氧胆酸钠和十二烷基硫酸钠提取的蛋白质混合物在质量上反映了膜的蛋白质组成。另一方面,在非离子型洗涤剂中,曲拉通X-100似乎主要能够提取一条带(很可能是带3区),而吐温80显然没有提取任何膜蛋白。详细描述了洗涤剂浓度、介质组成和实验程序。

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