Two-dimensional crystals of the Kdp-ATPase of Escherichia coli

Abstract

A variant form of the Kdp-ATPase of Escherichia coli mas overproduced to a level approaching 37% of the protein in the inner membrane of this organism. Membranes from overproducing cells were prepared with an inside-out orientation. Incubation of the membranes on ice for 1-2 weeks in the presence of sodium vanadate resulted in the formation of two-dimensional crystals of the Kdp-ATPase. The calculated projection map of the p1 crystal form showed three prominent density peaks at a resolution of 22 Angstrom. This technique is a useful and simple method to obtain low-resolution structures of membrane proteins.