Probing DNA Base-Dependent Leaving Group Kinetic Effects on the DNA Polymerase Transition State.

ABSTRACT: We examine the DNA polymerase β (pol β) transition state (TS) from a leaving group pre-steady-state kinetics perspective by measuring the rate of incorporation of dNTPs and corresponding novel β,γ-CXY-dNTP analogues, including individual β,γ-CHF and -CHCl diastereomers with defined stereochemistry at the bridging carbon, during the formation of right (R) and wrong (W) base pairs. Brønsted plots of log k_pol versus p K_a4 of the leaving group bisphosphonic acids are used to interrogate the effects of the base identity, the dNTP analogue leaving group basicity, and the precise configuration of the C-X atom in R and S stereoisomers on the rate-determining step ( k_pol). The dNTP analogues provide a range of leaving group basicity and steric properties by virtue o

SUBMITTER: Oertell K 

PROVIDER: S-EPMC6039227 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature