GENERATION OF PROTON-MOTIVE FORCE BY AN ARCHAEAL TERMINAL QUINOL OXIDASE FROM SULFOLOBUS-ACIDOCALDARIUS

The terminal quinol oxidase of the cytochrome aa(3) type was isolated from the extreme thermoacidophilic archaeon Sulfolobus acidocaldarius. In micellar solution, the enzyme oxidized various quinols and exerted the highest activity with the physiological substrate caldariella qui nol. The enzyme was functionally reconstituted into monolayer liposome s composed of archaeal tetraether lipids also derived from S. acidocal darius. With the electron donor system ascorbate and N,N,N',N'-tetrame thyl-p-phenylenediamine, the reconstituted enzyme was more active in t he archaeal lipids as compared to lipids derived from Escherichia coli at temperatures above 50 degrees C. Due to the low proton permeabilit y of the tetraether lipids, it was possible to generate a steady-state transmembrane electrical potential (Delta Psi, interior negative), an d transmembrane pH gradient (Delta pH, interior alkaline) at temperatu res up to 70 degrees C. The successful functional reconstitution of th e cytochrome aa(3)-type quinol oxidase from Sulfolobus identifies it a s the key energy converter in the respiratory system of this hyperther mophilic archaeon.