Detergent removal by non-polar polystyrene beads - Applications to membrane protein reconstitution and two-dimensional crystallization

被引:141
作者
Rigaud, JL
Levy, D
Mosser, G
Lambert, O
机构
[1] Inst Curie, Sect Rech, UMR CNRS 168, F-75231 Paris, France
[2] LRC CEA 8, F-75231 Paris, France
来源
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS | 1998年 / 27卷 / 04期
关键词
bio-beads; detergents; liposomes; proteoliposomes; reconstitution 2D crystallization; membrane proteins;
D O I
10.1007/s002490050138
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Detergent removal from lipid-protein-detergent micellar solutions is the most successful strategy for reconstitution of integral membrane proteins into proteoliposomes or into two-dimensional crystals. This review establishes the potential of polystyrene beads as a simple alternative to other conventional detergent removing strategies such as dialysis, gel chromatography and dilution. Kinetics and equilibrium aspects of removal of different detergents by hydrophobic adsorption onto polystyrene beads have been systematically investigated. A mechanism of adsorption onto polystyrene beads is proposed and provide useful information about the use of these beads in reconstitution experiments. The usefulness of this detergent removal strategy to produce quasi-ideal proteoliposomes is evaluated by considering the morphology and the size of the reconstituted vesicles, the homogeneity in size and protein distribution, the final protein orientation and the permeability of resulting proteoliposomes, Finally, the advantages of detergent removal by polystyrene beads as an alternative to conventional dialysis in two-dimensional crystallization trials are evaluated through review of recent structural reconstitution studies.
引用
收藏
页码:305 / 319
页数:15
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