Glycophorin A transmembrane domain dimerization in plasma membrane vesicles derived from CHO, HEK 293T, and A431 cells.
Glycophorin A transmembrane domain dimerization in plasma membrane vesicles derived from CHO, HEK 293T, and A431 cells.
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源自 CHO、HEK 293T 和 A431 细胞的质膜囊泡中血型糖蛋白 A 跨膜结构域二聚化。
DOI：
10.1016/j.bbamem.2013.03.022
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Hristova,Kalina
中科院分区：
文献类型：
--
作者：
Sarabipour,Sarvenaz;Hristova,Kalina
膜蛋白相互作用是生物功能的基础，发生在复杂的细胞膜环境中。质膜衍生的囊泡是一个模型系统，它允许研究膜蛋白之间的相互作用，而不需要提取、纯化和重组成脂双层。质膜囊泡可以由不同的细胞系和不同的方法产生，提供了丰富的、类似于自然的模型系统。然而，有了这些选择，就出现了不同类型的囊泡制剂如何影响膜蛋白之间的相互作用的问题。在这里，我们使用血糖蛋白A跨膜结构域(GPA)作为模型系统来解决这个问题。我们比较了来自中国仓鼠卵巢(CHO)、人胚胎肾293T(HEK 293T)和A431细胞的6种不同囊泡制剂中GPA的二聚化。我们通过使用基于FRET的方法来实现这一点，该方法产生FRET效率、供体浓度和每个囊泡中的受体浓度。我们发现，囊泡制备方法对GPA二聚化没有统计学意义的影响。基于这些结果，我们认为所研究的六种质膜制剂中的任何一种都可以作为研究膜蛋白相互作用的模型系统。
Membrane protein interactions, which underlie biological function, take place in the complex cellular membrane environment. Plasma membrane derived vesicles are a model system which allows the interactions between membrane proteins to be studied without the need for their extraction, purification, and reconstitution into lipid bilayers. Plasma membrane vesicles can be produced from different cell lines and by different methods, providing a rich variety of native-like model systems. With these choices, however, questions arise as to how the different types of vesicle preparations affect the interactions between membrane proteins. Here we address this question using the glycophorin A transmembrane domain (GpA) as a model system. We compare the dimerization of GpA in six different vesicle preparations derived from Chinese hamster ovary (CHO), Human Embryonic Kidney 293T (HEK 293T) and A431 cells. We accomplish this with the use of a FRET-based method which yields the FRET efficiency, the donor concentration, and the acceptor concentration in each vesicle. We show that the vesicle preparation protocol has no statistically significant effect on GpA dimerization. Based on these results, we propose that any of the six plasma membrane preparations investigated here can be used as a model system for studies of membrane protein interactions.