Figure 5.

Expression, purification, and enzyme activity of the PLCζ^H233L and PLCζ^H398P mutant proteins. (A) The affinity-purified NusA-fusion proteins for PLCζ^H233L and PLCζ^H398P (1 μg) analyzed by 7% SDS-PAGE (left panel) or by immunoblot analysis using anti-PLCζ polyclonal (V-37; 1:10,000 dilution; middle panel) or anti-NusA monoclonal antibody (1:20,000 dilution; right panel). (B) The [³H]PIP₂ hydrolysis activity of the purified PLCζ^H233L and PLCζ^H398P proteins, n = 3 ± standard error of the mean, determined using two different preparations of recombinant protein and with each experiment performed in duplicate.