UV-induced electron transfer between triethylamine and 5-bromo-2′-deoxyuridine. A puzzle concerning the photochemical debromination of labeled DNA
Paweł Wityk , Magdalena Zdrowowicz , Justyna Wiczk , Janusz Rak
Abstract5-Bromo-2′-deoxyuridine (BrdU) photosensitizes DNA to strand break formation. However, this type of photodamage is completely quenched by the presence of triethylamine (TEA) which originates from RP-HPLC purification commonly employed by oligonucleotide providers. While the presence of TEA in oligonucleotide samples does not interfere with PCR or other molecular biology applications, the mechanism of photochemical reaction proceeding in the labeled DNA is dramatically changed due to the photoinduced electron transfer (PET) between the photoexcited BrdU and the ground state TEA. For the first time, we demonstrated that the latter process produces 2′-deoxyuridne2′-deoxyuridine (debromination) in the labeled DNA instead of the expected strand break. PET between TEA and BrdU was additionally confirmed by the UV irradiations of aqueous solutions containing both species. Indeed, the efficient formation of 2′-deoxyuridine was observed in the studied photolytes. Moreover, we showed the formation of an additional product in these binary mixtures, i.e. imidazole derivative, that is not formed in DNA and was reported in the literature in the context of dark rather than photochemical processes. Using mass spectrometry we demonstrated that the amount of TEA impurity in the commercial samples of oligos exceeds up to 3 orders of magnitude that of the purchased DNA.
|Other language title versions|
|Journal series||Journal of Pharmaceutical and Biomedical Analysis, ISSN 0731-7085, (A 35 pkt)|
|Publication size in sheets||0.5|
|Keywords in English||triethylamine, photoinduced electron transfer, synthetic oligonucleotides, mass spectrometry, debromination, strand break|
|ASJC Classification||; ; ; ;|
|Score||= 35.0, 30-09-2019, ArticleFromJournal|
|Publication indicators||: 2017 = 1.179; : 2017 = 2.831 (2) - 2017=2.863 (5)|
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.