Structural analysis of ATP synthase from bovine heart mitochondria

This chapter discusses the structural analysis of Adenosine triphosphate (ATP) synthase from bovine heart mitochondria and the purification and crystallization of bovine F1-ATPase, the purification and characterization of monodisperse bovine F1F0-ATPase and F0, the bacterial expression and purification of the bovine ATPase inhibitor protein, and bovine ATPase and the reconstitution of the stalk and the F1 stalk complexes. The membrane association and the complexity of ATP synthase remain as obstacles that impede progress toward this objective. Pure monodisperse bovine F1F0-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 F1-ATPase and various subunits of F1F0-ATPase. © 1995, Elsevier Inc. All rights reserved.