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N 2 ‐Substituted 2′‐Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ
Author(s) -
Gowda A. S. Prakasha,
Lee Marietta,
Spratt Thomas E.
Publication year - 2017
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201611607
Subject(s) - dna polymerase , chemistry , polymerase , deoxyguanosine , microbiology and biotechnology , dna , substrate (aquarium) , in vivo , reactivity (psychology) , azide , stereochemistry , biochemistry , biology , organic chemistry , genetics , medicine , ecology , alternative medicine , pathology
N 2 ‐Alkyl‐2′‐deoxyguanosine triphosphate (N 2 ‐alkyl‐dGTP) derivatives with methyl, butyl, benzyl, or 4‐ethynylbenzyl substituents were prepared and tested as substrates for human DNA polymerases. N 2 ‐Benzyl‐dGTP was equal to dGTP as a substrate for DNA polymerase κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N 2 ‐4‐ethynylbenzyl‐dG with wild‐type and pol κ deficient mouse embryonic fibroblasts. CuAAC reaction with 5(6)‐FAM‐azide demonstrated that only cells containing pol κ were able to incorporate N 2 ‐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 κ in vivo.