FIGURE 5.

In vitro methyltransferase activities of purified His-Ste14p and variants. Purified protein (0.3 μg) was reconstituted in a 3000-fold excess of E. coli polar lipid by 20-fold rapid dilution in 100 mm MES, pH 7.0, and then incubated on ice for 10 min. Reactions contained the reconstituted purified proteins, 200 μm AFC, and 20 μm [¹⁴C]SAM and were incubated for 30 min at 30 °C. The reactions were terminated by the addition of 1 m NaOH, 1% SDS, and activity was quantified as described under “Experimental Procedures.” Each reaction was performed three times in duplicate, and error bars represent ±S.D. WT, wild type.