Figure 4.

ProNAc conjugation catalyzed by HhC using hairpin-forming steramer, S-dHP₃₉. (A) Kinetic analysis: activity of C−H−Y toward S-dHP₃₉ at the indicated concentrations, monitored by loss of FRET. (B) Michaelis−Menten: initial rate of reaction using FRET assay plotted as a function of increasing S-dHP₃₉. (C) Gel validation of ProNAc formation: reactions of C−H−Y in buffer only (NS), and decreasing concentrations of S-dHP₃₉. ProNAcs were detected using the DNA stain, GelRed (upper); total protein was visualized with Coomassie blue dye (lower). Calculated molecular weight of conjugate, CFP:S-dHP₃₉ is 38.8 kDa. Percent precursor consumed, %PC.