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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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Published in: | Radiation research 1994-05, Vol.138 (2), p.177-185 |
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Main Authors: | , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that cite this one |
Online Access: | Get full text |
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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. |
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ISSN: | 0033-7587 1938-5404 |
DOI: | 10.2307/3578587 |