Fig. 4. Binding of C1D and Translin to TRAX is mutually exclusive. C1D was expressed in bacteria and immobilized on Ni-NTA column as explained in the legend to Fig. 2. IVTT TRAX and Translin products were incubated with the immobilized C1D either alone (lanes 3 and 4) or together. TRAX (T) and Translin (t) IVTT products were incubated simultaneously (S) with the immobilized C1D for 2 hours before washing (lane 5), TRAX and Translin IVTT products were pre-incubated (PI) for 1 hour before adding to immobilized C1D and incubated for another hour (lane 6), IVTT product of TRAX was incubated with the immobilized C1D (C) for 1 hour and then Translin was added and incubated for an additional hour (lane 7). The plasmid expressing His only (pQE) was expressed in bacteria, purified and immobilized under the same conditions as C1D and was used in lane 8 as a negative control.
