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. Author manuscript; available in PMC: 2010 Jul 17.

Published in final edited form as: J Mol Biol. 2009 May 23;390(3):368–379. doi: 10.1016/j.jmb.2009.05.037

Concentration dependence on the rate of dTTP incorporation into 21-19/41A-mer (Table 1). A pre-incubated solution of enzyme (300 nM) and 5′-[³²P]-labeled 21-19/41A-mer DNA (30 nM) was mixed with increasing concentrations of dTTP for various times prior to be quenched by 0.37 M EDTA. The observed rate constants (k_obs) were plotted against the concentrations of dTTP and the data were fit to Equation 2 (Materials and Methods). (A) For fPolλ R386A, a k_p of 1.3 ± 0.1 s⁻¹ and a K_d of 15 ± 5 µM were determined, and (B) for fPolλ R386E, a k_p of 0.005 ± 0.001 s⁻¹ and a K_d of 1000 ± 410 µM were determined.

Concentration dependence on the rate of dTTP incorporation into 21-19/41A-mer (Table 1). A pre-incubated solution of enzyme (300 nM) and 5′-[³²P]-labeled 21-19/41A-mer DNA (30 nM) was mixed with increasing concentrations of dTTP for various times prior to be quenched by 0.37 M EDTA. The observed rate constants (k_obs) were plotted against the concentrations of dTTP and the data were fit to Equation 2 (Materials and Methods). (A) For fPolλ R386A, a k_p of 1.3 ± 0.1 s⁻¹ and a K_d of 15 ± 5 µM were determined, and (B) for fPolλ R386E, a k_p of 0.005 ± 0.001 s⁻¹ and a K_d of 1000 ± 410 µM were determined.