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. Author manuscript; available in PMC: 2006 Dec 28.
Published in final edited form as: J Biol Chem. 2005 Jan 10;280(13):12190–12200. doi: 10.1074/jbc.M412859200

Fig. 1.

Fig. 1

Active site titration of MuLV and HIV-1 RT. Pre-steady-state and steady-state kinetics of MuLV RT (A) and HIV-1 RT (B) incorporating correct dTTP onto the 32P-labeled 23-mer T primer annealed to the 38-mer RNA template were analyzed. Reactions were carried out at the indicated times by mixing together prebound RT (50–75 nm)·T/P (300 nm) with 800 μm dTTP under rapid quench conditions (see “Experimental Procedures”). The data were fit into the burst equation (Eq. 1) as indicated by the solid line, which provides a measure of the active concentration of RT (Amp), the observed first-order rate constant for the burst phase (kobs), and the first-order rate constant for the linear phase (k) for MuLV and HIV RT. On the matched T/P, the active concentration (Amp) of MuLV RT was 74.4 ± 0.9 nm. Its kobs was 28.95 ± 5.6 s−1, and its kss was 1.3 × 10−1 − 4.3 × 10−3 S−1. On matched T/P, the active concentration (Amp) of HIV-1 RT was 48.9 ± 4.7 nm. Its kobs was 129.25 ± 50.4 s−1 and its kss was 2.2 × 10−2 ± 1.6 × 10−2 S −1.