Molecular imaging of Escherichia coli F0F1‐ATPase in reconstituted membranes using atomic force microscopy

The structure of Escherichia coli F0F1‐ATPase (ATP synthase), and its F0 sector reconstituted in lipid membranes was analyzed using atomic force microscopy (AFM) by tapping‐mode operation. The majority of F0F1‐ATPases were visualized as spheres with a calculated diameter of , and a height of from the membrane surface. F0 sectors were visualized as two different ring‐like structures (one with a central mass and the other with a central hollow of depth) with a calculated outer diameter of . The two different images possibly represent the opposite orientations of the complex in the membranes. The ring‐like projections of both images suggest inherently asymmetric assemblies of the subunits in the F0 sector. Considering the stoichiometry of F0 subunits, the area of the image observed is large enough to accommodate all three F0 subunits in an asymmetric manner.

[1]  Y. Kagawa,et al.  Resolution of the membrane moiety of the H+-ATPase complex into two kinds of subunits. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[2]  R. Aggeler,et al.  Cryoelectron microscopy of Escherichia coli F1 adenosinetriphosphatase decorated with monoclonal antibodies to individual subunits of the complex. , 1989, Biochemistry.

[3]  Jan Pieter Abrahams,et al.  Structure at 2.8 Â resolution of F1-ATPase from bovine heart mitochondria , 1994, Nature.

[4]  P. Boyer,et al.  The binding change mechanism for ATP synthase--some probabilities and possibilities. , 1993, Biochimica et biophysica acta.

[5]  U. Lücken,et al.  Structure of the ATP synthase complex (ECF1F0) of Escherichia coli from cryoelectron microscopy. , 1990, Biochemistry.

[6]  M. Girvin,et al.  Hairpin folding of subunit c of F1Fo ATP synthase: 1H distance measurements to nitroxide-derivatized aspartyl-61. , 1994, Biochemistry.

[7]  R. Aggeler,et al.  Ligand-dependent structural variations in Escherichia coli F1 ATPase revealed by cryoelectron microscopy. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[8]  The Bacteria , 1881, Nature.

[9]  Y. Moriyama,et al.  One-step purification of Escherichia coli H(+)-ATPase (F0F1) and its reconstitution into liposomes with neurotransmitter transporters. , 1991, The Journal of biological chemistry.

[10]  T. Noumi,et al.  ATP synthase (H+-ATPase): results by combined biochemical and molecular biological approaches. , 1989, Annual review of biochemistry.

[11]  The F0 complex of the Escherichia coli ATP synthase. Investigation by electron spectroscopic imaging and immunoelectron microscopy. , 1995, European journal of biochemistry.

[12]  M. Girvin,et al.  Determination of local protein structure by spin label difference 2D NMR: the region neighboring Asp61 of subunit c of the F1F0 ATP synthase. , 1994, Biochemistry.

[13]  David Keller,et al.  Scanning force microscopy of nucleic acids and nucleoprotein assemblies , 1993 .

[14]  R. H. Fillingame CHAPTER 12 – Molecular Mechanics of ATP Synthesis by F1F0-Type H+ -Transporting ATP Synthases , 1990 .

[15]  K. Takeyasu,et al.  Molecular imaging of Na+,K+‐ATPase in purified kidney membranes , 1994, FEBS letters.