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Inhibition of Peroxyoxalate Chemiluminescence by Intercalation of Fluorescent Acceptors between DMA Bases
Author(s) -
Alba F. Javier,
Daban JoanRamon
Publication year - 1999
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.1751-1097.1999.tb03305.x
Subject(s) - peroxyoxalate , chemiluminescence , chemistry , ethidium bromide , intercalation (chemistry) , photochemistry , fluorescence , dimer , cationic polymerization , dna , inorganic chemistry , chromatography , organic chemistry , biochemistry , physics , quantum mechanics
— We have examined the ability of different fluorescent DNA dyes to become chemically excited by the peroxyoxalate chemiluminescent reaction. The intercalating dyes ethidium bromide and propidium iodide, and the bis ‐intercalating dyes ethidium homodimer‐1, benzoxazolium‐4‐pyridinium dimer‐1 and benzoxazolium‐4‐quinolinium dimer‐1, exhibit an intense Chemiluminescence when they are excited by the bis (2,4,6‐trichlorophenyl)oxalate (TCPO)‐H 2 O 2 reaction in the absence of DNA. However, the Chemiluminescence of these dyes is very low when they are bound to double‐stranded DNA (dsDNA). In contrast, the minor groove‐binding dye Hoechst 33258 excited by the TCPO‐H 2 O 2 reaction shows approximately the same Chemiluminescence intensity when it is free in solution or complexed with dsDNA. Structural alterations or partial dissociation of dsDNA‐ bis ‐intercalating dye complexes produced by the addition of acetone, NaCl, MgCl 2 or the cationic surfactant cetyltrimethylammonium bromide increases the Chemiluminescence intensity. A moderate Chemiluminescence intensity is observed when bis ‐intercalating dyes are complexed with single‐stranded DNA. Our results indicate that the energy from the intermediates produced in the peroxyoxalate chemiluminescent reaction cannot be efficiently transferred to fluorescent dyes complexed with DNA; chemiexcitation is almost completely inhibited when dyes are buried in the dsDNA structure by intercalation between the base pairs.