Isolation, characterization and structure of bacterial flagellar motors containing the switch complex.

A putative complex of the three switch proteins, FliG, FliM and FliN appears to be directly involved in torque generation and control of direction of rotation. We have developed a preparative procedure for flagellar motors that retains these proteins as evidenced by Western blots using anti-FliG, anti-FliM and anti-FliN antibodies. Immunogold labeling with these three antibodies shows that the three switch proteins are localized to the motor. Electron micrographs of frozen-hydrated preparations reveal a large, new component we have termed the "C ring complex" attached to the cytoplasmic face of the M ring. In a three-dimensional reconstruction of the cylindrically averaged structure, the M-S ring complex appears thicker and wider by the addition of extra material to the cytoplasmic surface of the M ring. In addition, extending into the cytoplasm from the thickened M ring is the C ring complex, a thin-walled cylinder having a length of 170 A and an outer diameter of 450 A compared to the 290 A diameter of the M ring. We provide evidence that the thickened M ring contains FliG and that the C ring complex may contain FliM and FliN. The large diameter of the C ring complex may permit interaction with the M ring and with the circlet of studs thought to be the MotA/MotB complex.