Quantification of radiation-induced hydroxyl radicals within nucleohistones using a molecular fluorescent probe

G. M. Makrigiorgos, M. Folkard, C. Huang, E. Bump, J. Baranowska-Kortylewicz, S. K. Sahu, B. D. Michael, A. I. Kassis

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16 Scopus citations

Abstract

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 in neutral aqueous solutions. It is shown that (a) the fluorescent product 7-OH-SECCA cannot be generated by direct radiation effects after γ or neutron irradiation of SECCA; (b) when SECCA-labeled histone is complexed with DNA to form nucleohistone, the physical properties of the modified nucleohistone are similar to those of unlabeled nucleoprotein; and (c) after irradiation of SECCA-labeled nucleohistone, a linear induction of the fluorescence signal is observed within the radiation doses examined (0.3-30 Gy). Since the sample remains available for further studies after registration of the optical signal, the current approach should permit the investigator to correlate in a single sample the localization and frequency of ·OH formation with the results of other assays.

Original languageEnglish (US)
Pages (from-to)177-185
Number of pages9
JournalRadiation Research
Volume138
Issue number2
DOIs
StatePublished - 1994

ASJC Scopus subject areas

  • Biophysics
  • Radiation
  • Radiology Nuclear Medicine and imaging

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    Makrigiorgos, G. M., Folkard, M., Huang, C., Bump, E., Baranowska-Kortylewicz, J., Sahu, S. K., Michael, B. D., & Kassis, A. I. (1994). Quantification of radiation-induced hydroxyl radicals within nucleohistones using a molecular fluorescent probe. Radiation Research, 138(2), 177-185. https://doi.org/10.2307/3578587