Abstract
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 κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N2-4-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 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 κ in vivo.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2628-2631 |
| Number of pages | 4 |
| Journal | Angewandte Chemie - International Edition |
| Volume | 56 |
| Issue number | 10 |
| DOIs | |
| State | Published - Jan 1 2017 |
Fingerprint
All Science Journal Classification (ASJC) codes
- Catalysis
- Chemistry(all)
Cite this
}
N2-Substituted 2′-Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ. / Gowda, A. S.Prakasha; Lee, Marietta; Spratt, Thomas E.
In: Angewandte Chemie - International Edition, Vol. 56, No. 10, 01.01.2017, p. 2628-2631.Research output: Contribution to journal › Article
TY - JOUR
T1 - N2-Substituted 2′-Deoxyguanosine Triphosphate Derivatives as Selective Substrates for Human DNA Polymerase κ
AU - Gowda, A. S.Prakasha
AU - Lee, Marietta
AU - Spratt, Thomas E.
PY - 2017/1/1
Y1 - 2017/1/1
N2 - 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 κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N2-4-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 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 κ in vivo.
AB - 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 κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N2-4-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 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 κ in vivo.
UR - http://www.scopus.com/inward/record.url?scp=85011312045&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85011312045&partnerID=8YFLogxK
U2 - 10.1002/anie.201611607
DO - 10.1002/anie.201611607
M3 - Article
C2 - 28140505
AN - SCOPUS:85011312045
VL - 56
SP - 2628
EP - 2631
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
SN - 1433-7851
IS - 10
ER -