Fig. 5.
Determination of critical concentration of toluene to permeabi-lize cells. E. coli cells expressing PheP with a single cysteine in periplas-mic domain P5 [5] were incubated for 5 min at room temperature in the presence of varying concentrations of toluene prior to reaction with 100 µM of MPEOB. The cells were lysed and solubilized membrane proteins were subjected to immunoprecipitation with anti-PheP polyclonal antibody. The immunoprecipitates were subjected to SDS-PAGE and biotinylated proteins were detected using avidin-HRP and chemiluminescence.