FIGURE 6.

Purification of recombinant wild-type and mutant Mtr4p. 6xHis-tagged Mtr4p was expressed in E. coli and purified using TALON metal affinity resin and anion exchange chromatography (Materials and Methods). Equal amounts of purified protein, as determined by Bradford assay, were resolved on 4%–12% gradient SDS-PAGE gel. (A) Silver staining and (B) Western blot with anti-His antibody were used to visualize and quantify the amount of purified protein in each preparation. (Lanes 1,4) WT Mtr4p; (lanes 2,5) Mtr4-20p; (lanes 3,6) Mtr4-21p contains the changes DEvH to AAvH.