0000000000889934
AUTHOR
K. Zwicker
111 Bioluminescence analysis and numerical evaluation of ATP-synthesis by native and reconstituted membranes containing bacterial ATP-synthase
ATP-synthase is a large membrane protein complex, which plays a key role in the energy metabolism of most organisms. It consists of at least eight types of subunits and can be isolated and purified from several organisms, e.g. bacteria. The enzyme couples two reversible reactions: vectorial proton transport through a membrane and synthesis of the energy rich molecule ATP. Both can be investigated with vesicles from native membranes or with reconstituted liposomes from purified ATPsynthase. The analysis is complicated because ATP-synthase catalyzes ATP-synthesis as well as ATP-hydrolysis. Furthermore the ATP level of membrane samples is influenced by adenylate kinase activities of other enzy…
Reconstitution of bacteriorhodopsin and ATP synthase from Micrococcus luteus into liposomes of the purified main tetraether lipid from Thermoplasma acidophilum: proton conductance and light-driven ATP synthesis.
The archaebacterium Thermoplasma acidophilum is cultivated at 59 degrees C in a medium containing sulfuric acid of pH 2. The purified bipolar membrane spanning main phospholipid (MPL) of this organism can be used to produce stable liposomes of 100-500 nm in diameter either using a French pressure cell detergent dialysis or sonication. Despite a potassium diffusion potential of 186 mV very low ionic permeability of sonicated MPL liposomes was measured using the potassium binding fluorescent indicator benzofuran isophthalate PBF1, which measures net K+ uptake. The latter also remained very low, in the presence of the K(+) ionophore valinomycin and palmitic acid. Addition of valinomycin and th…