N2 -substituted 2'-deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase Kappa
Biochemistry and Molecular Biology
N2 -Alkyl-2'-deoxyguanosine triphosphate (N2 -alkyl-dGTP) derivatives with methyl, butyl, benzyl, or 4-ethynylbenzyl substituents were prepared and tested as substrates for human DNA polymerases. N2 -Benzyl-dGTP was equal to dGTP as a substrate for DNA polymerase kappa (pol kappa), but was a poor substrate for pols beta, delta, eta, iota, or nu. In vivo reactivity was evaluated through incubation of N2 -4-ethynylbenzyl-dG with wild-type and pol kappa deficient mouse embryonic fibroblasts. CuAAC reaction with 5(6)-FAM-azide demonstrated that only cells containing pol kappa were able to incorporate N2 -4-ethynylbenzyl-dG into the nucleus. This is the first instance of a Y-family-polymerase-specific dNTP, and this method could be used to probe the activity of pol kappa in vivo.
Gowda, A., Lee, M., & Spratt, T. (2017). N2 -substituted 2'-deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase Kappa. Angewandte Chemie (International ed.in English), 56 (10), 2628-2631. https://doi.org/10.1002/anie.201611607