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3‐CARBETHOXYPSORALEN‐DNA PHOTOLESIONS: IDENTIFICATION AND QUANTITATIVE DETECTION IN YEAST AND MAMMALIAN CELLS OF THE TWO cis‐syn DIASTEREOISOMERS FORMED WITH THYMIDINE
Author(s) -
Moysan A.,
Vigny P.,
Dardalhon M.,
Averbeck D.,
Voituriez L.,
Cadet J.
Publication year - 1988
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1988.tb01663.x
Subject(s) - diastereomer , dna , yeast , chinese hamster , thymidine , chemistry , fluorescence , biochemistry , microbiology and biotechnology , stereochemistry , biology , physics , quantum mechanics
Abstract A chemical method for the identification and the quantitative detection of psoralen DNA furan‐side photoadducts formed in cells is presented. It is based on an enzymatic digestion of the purified DNA extracted from the treated cells and a further separation by high performance liquid chromatography of the modified nucleosides coupled to a highly sensitive fluorescence analysis and detection. Using this method, 3‐carbethoxypsoralen‐ DNA photoadducts formed in yeast and mammalian cells have been identified and quantified. The predominant photoadducts induced have been identified as two cis‐syn dThd( 5 6 4' 5' )3‐CPs diastereoisomers. In Chinese Hamster V79 cells treated with 3‐carbethoxypsoralen at 50 μM and irradiated at 365 nm with an incident dose of 24 kJ/m 2 , the two monoadducts could be quantitatively assessed at levels as low as 1.3 and 0.7 per 10 000 base pairs.