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. 2012 Sep 19;287(45):38090–38100. doi: 10.1074/jbc.M112.404954

FIGURE 3.

FIGURE 3.

Histidine mutations at Trp-102β and Trp-106β enhance FTase reactivity with dns-GCVDS. a, dependence of activity on the peptide concentration catalyzed by WT (circle) and W102H/W106H (square) FTase is shown. Reactivity was measured with purified enzymes using the fluorescence-based assay as described under “Experimental Procedures.” b, shown is reactivity of W102H, W106H, and W102H/W106H FTases with dns-GCVDS (solid bar) and dns-GCVAS (checkered bar) relative to WT FTase. Relative reactivity was calculated as (kcat/Km)variant/(kcat/Km)WT with kcat/Km measured as described under “Experimental Procedures.” The measured kcat/Km values are 41 ± 5 and 15,800 ± 130 m−1 s−1 for dns-GCVDS and dns-GCVAS, respectively (WT FTase), 430 ± 30 and 15,700 ± 100 m−1 s−1 (W102H FTase), 160 ± 20 and 8,900 ± 50 m−1 s−1 (W106H FTase), and 1,200 ± 100 and 9,000 ± 50 m−1 s−1 (W102H/W106H FTase).