Partial separation of inwardly pumping and outwardly pumping bacteriorhodopsin reconstituted liposomes by gel filtration

FEBS Letters

Rigaud, Jean-Louis¹  Seigneuret, Michel¹ 

[1] Service de Biophysique, Département de Biologie, CEN-Saclay, 91191 Gif-sur-Yvette Cedex, France

关键词: Bacteriorhodopsin;    Reconstitution;    Protein orientation;    Liposome size;    Gel filtration;    Pyranine;    BR;    bacteriorhodopsin;    Pipes;    1;    4-piperazine ethane sulfonate;    pyranine;    8-hydroxy-1;    3;    6-pyrenetrisulfonic acid;   

DOI  :  10.1016/0014-5793(88)80589-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Reconstitution of bacteriorhodopsin using the reverse-phase evaporation technique leads to the formation of both inwardly proton pumping and outwardly proton pumping liposomes [(1985) FEBS Lett. 188, 101–1061]. The reconstituted preparation was size fractionated using gel filtration chromatography on Sephacryl S1000. The direction of proton pumping was assessed in various fractions using the pH sensitive fluorescence probe pyranine. The proportion of outwardly pumping liposomes was shown to increase with decreasing liposome size. The larger liposomes (diameter larger than 200 nm) were shown to be pure inwardly pumping liposomes with almost homogeneous bacteriorhodopsin orientation.

【 授权许可】

Unknown