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Quantification of Radiation-Induced Hydroxyl Radicals within Nucleohistones Using a Molecular Fluorescent Probe

We present a method that specifically records ^{\bullet}{\rm OH}$ formation within histones and possibly at other sites in irradiated nucleohistone. The approach uses the radiation-induced fluorescence emissions from a chromatin-conjugated ^{\bullet}{\rm OH}$ detector, SECCA (a succinylated derivati...

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Bibliographic Details
Published in:Radiation research 1994-05, Vol.138 (2), p.177-185
Main Authors: Makrigiorgos, G. M., Folkard, M., Huang, C., Bump, E., Baranowska-Kortylewicz, J., Sahu, S. K., Michael, B. D., Kassis, A. I.
Format: Article
Language:English
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Summary:We present a method that specifically records ^{\bullet}{\rm OH}$ formation within histones and possibly at other sites in irradiated nucleohistone. The approach uses the radiation-induced fluorescence emissions from a chromatin-conjugated ^{\bullet}{\rm OH}$ detector, SECCA (a succinylated derivative of coumarin), that is converted to a fluorescent derivative, 7-hydroxy-SECCA (7-OH-SECCA), after interaction with ^{\bullet}{\rm 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 ^{\bullet}{\rm OH}$ formation with the results of other assays.
ISSN:0033-7587
1938-5404
DOI:10.2307/3578587