A fluorimetric method for the detection of copper-mediated hydroxyl free radicals in the immediate proximity of DNA.
AuthorsMakrigiorgos GM, Bump E, Huang C, Baranowska-Kortylewicz J, Kassis AI
JournalFree Radic Biol Med
PubMed ID7750791
'An optical method to detect copper-mediated hydroxyl free radicals generated close to DNA and other biomolecules has been developed. Low-molecular-weight polylysines were labeled with SECCA, a derivative of coumarin that generates the fluorescent 7-OH-SECCA following its interaction with hydroxyl free radicals in aqueous solution. These polylysines were then complexed with ... More
A method for detection of hydroxyl radicals in the vicinity of biomolecules using radiation-induced fluorescence of coumarin.
AuthorsMakrigiorgos GM, Baranowska-Kortylewicz J, Bump E, Sahu SK, Berman RM, Kassis AI
JournalInt J Radiat Biol
PubMed ID8096857
'A novel method is described to quantitate radiation-induced hydroxyl radicals in the vicinity of biomolecules in aqueous solutions. Coumarin-3-carboxylic acid (CCA) is a non-fluorescent molecule that, upon interaction with radiation in aqueous solution, produces fluorescent products. CCA was derivatized to its succinimidyl ester (SECCA) and coupled to free primary amines ... More
Measurement of hydroxyl radicals catalyzed in the immediate vicinity of DNA by metal-bleomycin complexes.
AuthorsChakrabarti S, Makrigiorgos GM, O'Brien K, Bump E, Kassis AI
JournalFree Radic Biol Med
PubMed ID8728024
'A recently developed sensitive fluorimetric assay has been used to examine whether free hydroxyl radicals (HO.) are generated in the immediate vicinity of DNA by Fe(II)-bleomycin. When aqueous solutions of SECCA (the succinimidyl ester of coumarin-3-carboxylic acid) are irradiated with gamma rays or incubated with Fe(II)-bleomycin or Fe (II)-EDTA in ... More
Quantification of radiation-induced hydroxyl radicals within nucleohistones using a molecular fluorescent probe.
AuthorsMakrigiorgos GM, Folkard M, Huang C, Bump E, Baranowska-Kortylewicz J, Sahu SK, Michael BD, Kassis AI
JournalRadiat Res
PubMed ID8183987
We present a method that specifically records .OH formation within histones and possibly at other sites in irradiated nucleohistone. The approach uses the radiation-induced fluorescence emissions from a chromatin-conjugated .OH detector, SECCA (a succinylated derivative of coumarin), that is converted to a fluorescent derivative, 7-hydroxy-SECCA (7-OH-SECCA), after interaction with .OH ... More
Accessibility of nucleic acid-complexed biomolecules to hydroxyl radicals correlates with their conformation: a fluorescence polarization spectroscopy study.
AuthorsMakrigiorgos GM, Bump E, Huang C, Baranowska-Kortylewicz J, Kassis AI
JournalInt J Radiat Biol
PubMed ID7930827
A fluorescence methodology has been developed to examine the relationship between the conformational state of specific biomolecules in simple chromatin models and their accessibility to hydroxyl radicals (OH). Polylysine and histone H1 were labelled with SECCA, the succinimidyl ester of coumarin-3-carboxylic acid, which generates the fluorescent derivative 7-OH-SECCA following its ... More
Generation of hydroxyl radicals by nucleohistone-bound metal-adriamycin complexes.
AuthorsChakrabarti S, Mahmood A, Kassis AI, Bump EA, Jones AG, Makrigiorgos GM
JournalFree Radic Res
PubMed ID8889487
A recently developed method has been utilized to demonstrate the generation of hydroxyl radicals (HO.) in the immediate proximity of DNA by copper(II)/iron(III)-adriamycin in the presence of ascorbate and hydrogen peroxide. SECCA, a succinylated derivative of coumarin, generates the fluorescent 7-hydroxy-SECCA following reaction with HO.. SECCA was coupled to polylysine ... More