[11] Structural analysis of ATP synthase from bovine heart mitochondria

Publisher Summary This chapter discusses the structural analysis of Adenosine triphosphate (ATP) synthase from bovine heart mitochondria and the purification and crystallization of bovine F 1 -ATPase, the purification and characterization of monodisperse bovine F 1 F 0 -ATPase and F 0 , the bacterial expression and purification of the bovine ATPase inhibitor protein, and bovine ATPase and the reconstitution of the stalk and the F 1 stalk complexes. The membrane association and the complexity of ATP synthase remain as obstacles that impede progress toward this objective. Pure monodisperse bovine F 1 F 0 -ATPase can be prepared from heart mitochondria as a product of a multi-enzyme preparation based on chromatography for the isolation of respiratory enzymes from mitochondria. Alternatively, it can be prepared from mitochondrial membranes by extraction with dodecyl-β-d-maltoside and chromatography on Diethylaminoethyl (DEAE) cellulose. The chapter concludes with the discussion on the interactions between F 1 -ATPase and various subunits of F 1 F 0 -ATPase.