Figure 4.

ClpX chaperone monomerizes TrfA in an ATP-dependent reaction. Column gel exclusion chromatography using Bio-Gel P-100 was performed as described in Materials and Methods after incubation of ³⁵S-radiolabeled wild-type His6-TrfA with or without ClpX chaperone (1.5 μg). After column separation, the amount of radioactivity was determined for each fraction. TrfA protein profiles after incubation in the presence of ClpX and ATP (2 mM) (▪); ClpX and ATPγS (2 mM) (□); and ATP (2 mM) alone (○). The positions of peak fractions of the molecular weight markers are indicated: BSA (66 kDa), ovalbumin (OA) (44 kDa), and carbonic anhydrase (CA) (29 kDa). Arrows, positions of the TrfA dimer and monomer forms.