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[用于揭示A431细胞中α-辅肌动蛋白1和α-辅肌动蛋白4的亚细胞分级分离方法的比较分析]

[The comparative analysis of subcellular fractionation methods for revealing of alpha-actinin 1 and alpha-actinin 4 in A431 cells].

作者信息

Bol'shakova A V, Petukhova O A, Pinaev G P, Magnusson K E

出版信息

Tsitologiia. 2009;51(2):122-9.

Abstract

Alpha-actinin 1 and alpha-actinin 4 belong to a family of actin-binding proteins with shared structural function and regulation of several processes in a cell. Based on previous data on different distribution of these proteins in the nucleus and cytoplasm, we have explored in detail the distribution of alpha-actinin 1 and alpha-actinin 4 in subcellular fractions in A431 cells spread on fibronectin. Several methods of subcellular fractionation were used. Complex approach allowed resuming that revealing of alpha-actinin isoforms in fractions depended on the composition of lysis buffer and preliminary low-temperature freezing of the cells. We have drawn a conclusion that alpha-actinin 4 can be found in all cytoplasmic and nuclear subfractions, while alpha-actinin 1 is characterized by cytoplasmic and membrane localization with specificity of its distribution tightly to the nuclear membrane.

摘要

α-辅肌动蛋白1和α-辅肌动蛋白4属于肌动蛋白结合蛋白家族,它们在细胞中具有共同的结构功能并调节多种过程。基于先前关于这些蛋白质在细胞核和细胞质中不同分布的数据,我们详细研究了铺展在纤连蛋白上的A431细胞亚细胞组分中α-辅肌动蛋白1和α-辅肌动蛋白4的分布。使用了几种亚细胞分级分离方法。综合方法表明,各组分中α-辅肌动蛋白同工型的显示取决于裂解缓冲液的组成以及细胞的初步低温冷冻。我们得出的结论是,在所有细胞质和细胞核亚组分中都能发现α-辅肌动蛋白4,而α-辅肌动蛋白1的特征是定位于细胞质和细胞膜,其分布特异性地紧密靠近核膜。

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