Use of the fluorescent pH probe pyranine to detect heterogeneous directions of proton movement in bacteriorhodopsin reconstituted large liposomes

FEBS Letters

Rigaud, Jean-Louis¹  Seigneuret, Michel¹ 

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

关键词: Bacteriorhodopsin;    Reconstitution;    Protein orientation;    Proton pumping;    Pyranine;    pH;    transmembrane pH gradient;    Pipes;    1;    4-piperazineethanesulfonic acid;   

DOI  :  10.1016/0014-5793(85)80883-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Light-induced proton pumping was measured in bacteriorhodopsin reconstituted large proteoliposomes (prepared by reverse-phase evaporation) using the fluorescence of internally trapped pyranine. From the effect of the one-sided inhibitor La³⁺, the presence of both inside-out and right-side-out oriented bacterio-rhodopsin was demonstrated. The use of pyranine at acidic pH allowed us to detect two distinct populations of proteoliposomes which exhibited oppositely directed proton pumping. This suggests that the two orientations, of bacteriorhodopsin are at least partially distributed in different proteoliposomes.

【 授权许可】

Unknown