Figure 5. Site-directed mutagenesis studies on PALcc.
PEAK-Rapid cells were transiently transfected with vectors encoding wild-type PALcc or each of the mutants indicated. After 24 hours, cells were rinsed and incubated in complete serum-free medium (m) and cells (c) were harvested in TES-mannitol-Triton buffer with protease inhibitors. Samples were analyzed by Western blot (0.8% of the medium; 4% of the cell extract) using an anti-peptide polyclonal antibody and aliquots were assayed for PAL activity. The activity of each sample is shown at the top of each gel lane.