Detecting proton flux across chromatophores driven by F0F1-ATPase using N-(fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt.

摘要: Abstract N -(Fluorescein-5-thiocarbamoyl)-1,2-dihexadecanoyl- sn -glycero-3-phosphoethanolamine, triethylammonium salt (F-DHPE) is a lipid fluorescence dye sensitive to pH changes and used in this study for detecting proton flux through F 0 1 -ATPase within chromatophores driven by ATP hydrolysis. F-DHPE easily labeled the outer surface of chromatophores. In range 7.0 9.0, intensity changes. The sensitivity especially great 8.2 so 8.6 was chosen as appropriate experimental condition. It shown that added not only acts quencher but also can be hydrolyzed pump protons into chromatophores, resulting restoration. A stimulator (NaSO 3 ) various types inhibitors (NaN , 5′-adenylyl imidodiphosphate [AMP–PNP], ′-dicyclohexylcarbodiimide [DCCD]) confirmed restoration caused ATP-driven flux. When loaded with one antibody (anti-β antibody) or two antibodies sheep rabbit second antibody), exhibits lower pumping activities, indicated possible mechanism inhibition on activity discussed.