Use of the pH sensitive fluorescence probe pyranine to monitor internal pH changes in Escherichia coli membrane vesicles

FEBS Letters

Damiano, E.¹  Leblanc, G.¹  Bassilana, M.¹  Rigaud, J.-L.² 

[1] Laboratoire Jean Maetz, Département de Biologie du CEA, 06230 Villefranche-sur-Mer, France;Service de Biophysique, Département de Biologie du CEA, 06230 Villefranche-sur-Mer, France

关键词: Escherichia coli;    Membrane vesicle;    Internal pH;    Pyranine;    Fluorescent pH probe;    Δψ and ΔpH;    electrical and chemical proton gradients;    Δ acetate;    transmembrane acetate concentration gradient;    TPP+ Br−;    tetraphenylphosphonium bromide;    PMS;    phenazine methosulfate;    FCCP;    carbonyl cyanide p-trifluoromethoxyphenylhydrazone;   

DOI  :  10.1016/0014-5793(84)80056-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Measurements of the fluorescent properties of 8-hydroxy-1,3,6-pyrenetrisulfonate (pyranine) enclosed within the internal space of Escherichia coli membrane vesicles enable recordings and quantitative analysis of: (i) changes in intravesicular pH taking place during oxidation of electron donors by the membrane respiratory chain; (ii) transient alkalization of the internal aqueous space resulting from the creation of outwardly directed acetate diffusion gradients across the vesicular membrane. Quantitation of the fluorescence variations recorded during the creation of transmembrane acetate gradients shows a close correspondence between the measured shifts in internal pH value and those expected from the amplitude of the imposed acetate gradients.

【 授权许可】

Unknown