Electron microscopy together with image analysis has been used to study the
structure of the intact F1F0-ATPsynthase from Escherichia coli. A procedur
e has been developed which allows preparation of detergent-free enzyme. Asi
de from the well known two-domain structure, images of F1F0 prepared by thi
s procedure show a number of additional features, including a second stalk,
which can be seen extending all the way from the F-0 to the top of the F-1
in some images, and a small protein on the very top of the F-1, which has
been identified as the delta subunit by decoration with a monoclonal antibo
dy. In light of these results, a refined model of the subunit arrangement o
f the complex is presented.