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

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

Research output: Contribution to journalArticle

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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 - May 27 1994

Fingerprint

Molecular Probes
hydroxyl radicals
Fluorescent Dyes
histones
Hydroxyl Radical
Histones
probes (equipment)
irradiation
chemical derivatives
Fluorescence
fluorescence
Radiation
Coumarins
chromatin
nucleoproteins
Nucleoproteins
probes
coumarin
Radiation Effects
Neutrons

ASJC Scopus subject areas

  • Biophysics
  • Radiation
  • Radiology Nuclear Medicine and imaging

Cite this

Quantification of radiation-induced hydroxyl radicals within nucleohistones using a molecular fluorescent probe. / Makrigiorgos, G. M.; Folkard, M.; Huang, C.; Bump, E.; Baranowska-Kortylewicz, Janina; Sahu, S. K.; Michael, B. D.; Kassis, A. I.

In: Radiation Research, Vol. 138, No. 2, 27.05.1994, p. 177-185.

Research output: Contribution to journalArticle

Makrigiorgos, GM, Folkard, M, Huang, C, Bump, E, Baranowska-Kortylewicz, J, Sahu, SK, Michael, BD & Kassis, AI 1994, 'Quantification of radiation-induced hydroxyl radicals within nucleohistones using a molecular fluorescent probe', Radiation Research, vol. 138, no. 2, pp. 177-185. https://doi.org/10.2307/3578587
Makrigiorgos, G. M. ; Folkard, M. ; Huang, C. ; Bump, E. ; Baranowska-Kortylewicz, Janina ; Sahu, S. K. ; Michael, B. D. ; Kassis, A. I. / Quantification of radiation-induced hydroxyl radicals within nucleohistones using a molecular fluorescent probe. In: Radiation Research. 1994 ; Vol. 138, No. 2. pp. 177-185.
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