Dye-ligand chromatographic purification of intact multisubunit membrane protein complexes: application to the chloroplast H+-F0F1-ATP synthase

n-Dodecyl-beta-D-maltoside was used as a detergent to solubilize the ammoni um sulphate precipitate of chloroplast F0F1-ATP synthase, which was purifie d further by dye-ligand chromatography. Upon reconstitution of the purified protein complex into phosphatidylcholine/phosphatidic acid liposomes, ATP synthesis, driven by an artificial Delta pH/Delta psi was observed. The hig hest activity was achieved with ATP synthase solubilized in n-dodecyl-beta- D-maltoside followed by chromatography with Red 120 dye. The optimal dye fo r purification with CHAPS was Green 5. All known subunits were present in t he monodisperse proton-translocating ATP synthase preparation obtained from chloroplasts.