Figure 9.
Reversed-phase HPLC analysis with fluorescence detection of reactions between N-dansyl-GCVIA and FPP (1) or dihydroester 4 catalyzed by ScPFTase. (A) N-dansyl-GCVIA (2.4 μM) incubated with FPP (1, 10 μM) and ScPFTase (24 nM). Chromatogram 1 (bottom): Reaction before the addition of the enzyme with the peptide eluting at tR=25.7 min. Chromatogram 2 (top): Reaction after 5 min with the prenylated peptide (11) appearing at tR=41.5 min. (B) N-dansyl-GCVIA (2.4 μM) incubated with dihydroester (4, 10 μM) and ScPFTase (24 nM). Chromatogram 1 (bottom): Reaction before the addition of the enzyme with the peptide substrate at tR=25.7 min. Chromatogram 2 (top): Reaction after 23 h with the prenylated peptide (14) appearing at tR=36.0 min and some starting material still present.