期刊
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
卷 1798, 期 11, 页码 2094-2101出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbamem.2010.07.034
关键词
Multidrug resistance; ABCG2; Functionalized lipid layer; Electron microscopy; Sf9 insect cells; Flp-ln-293 cells
资金
- Institut Curie
- CNRS
- Universite Lyon 1 [UMR 5086]
- Agence Nationale de la Recherche [ANR-06-BLAN-0420, ANR-PCV06-135269, ANR-PCV06-135175]
- Ligue Nationale contre le Cancer
- Agence Nationale de la Recherche (ANR) [ANR-06-BLAN-0420] Funding Source: Agence Nationale de la Recherche (ANR)
ABCG2 is a human membrane ATP-binding cassette half-transporter that hydrolyzes ATP to efflux a large number of chemotherapeutic agents. Several oligomeric states of ABCG2 from homodimers to dodecamers have been reported depending on the overexpression systems and/or the protocols used for purification. Here, we compared the oligomeric state of His(6)-ABCG2 expressed in Sf9 insect cells and in human Flp-In-293/ABCG2 cells after solubilization in mild detergents. His(6)-ABCG2 was purified through a new approach involving its specific recognition onto a functionalized lipid layer containing a Ni-NTA lipid. This approach allowed the purification of His-ABCG2 in presence of all solubilized membrane components that might be involved in the stabilisation of native oligomers and without requiring any additional washing or concentration passages. ABCG2 purified onto the NiNTA lipid surfaces were directly analyzed by electron microscopy and by biochemical assays. Altogether, our data are consistent with a tetrameric organization of ABCG2 when expressed in either heterologous Sf9 insect cells or in human homologous cells. (C) 2010 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据