Identification of torque generation part in flagellar motor by using fluorescence polarization

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The microscopic system to detect the polarization of fluorescent molecules was develoved. By using this microscopic system, the polarization of fluorescence from a single GFP molecule was detected, and the direction of a GFP molecule on the surface of coverslip was estimated from the fluorescent intensities of GFP in P-polarization and S-polarization. The microscopic system to simultaneously observe the fluorescence from GFP-fusion proteins and the rotation of flagellar motor was also constructed. By using this microscopic system, it was demonstrated that the rotational switch of a motor was directly regulated by the binding and dissociation of phosphorylated CheY by simultaneously visualizing CheY tagged with GFP and the rotational switching of a motor in live cells. The results of this study was published from Sci. Signal. 2014. 7: ra32.