A method for detection of hydroxyl radicals in the vicinity of biomolecules using radiation-induced fluorescence of coumarin

G. M. Makrigiorgos, J. Baranowska-Kortylewicz, E. Bump, S. K. Sahu, R. M. Berman, A. I. Kassis

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

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 of albumin, avidin, histone-H1, polylysine, and an oligonucleotide. When SECCA-biomolecule conjugates were irradiated, the relationship between induced fluorescence and dose was linear in the dose range examined (0.01-10 Gy). The fluorescence excitation spectrum of irradiated SECCA-biomolecule conjugates was very similar to that of 7-hydroxy-SECCA-biomolecule conjugates, indicating the conversion of SECCA to 7-hydroxy-SECCA following irradiation. Control studies in environments that excluded certain radiation-induced water radicals for both the conjugated and unconjugated forms of irradiated SECCA demonstrated that: (1) the induction of fluorescence is mediated by the hydroxyl radical; (2) the presence of oxygen enhances induced fluorescence by a factor of about 1.4, and (3) other primary water radicals and secondary radicals caused by interaction of primary water radicals with biomolecules do not significantly influence the induced fluorescence. The data indicate that the induction of fluorescence on SECCA-biomolecule conjugates records specifically the presence of the hydroxyl radical in the immediate vicinity of the irradiated biomolecule. The method is rapid and sensitive, uses standard instrumentation, and the sample remains available for further studies.

Original languageEnglish (US)
Pages (from-to)445-458
Number of pages14
JournalInternational Journal of Radiation Biology
Volume63
Issue number4
DOIs
StatePublished - Jan 1 1993

Fingerprint

coumarin
hydroxyl radicals
Biomolecules
Hydroxyl Radical
Fluorescence
fluorescence
Radiation
radiation
carboxylic acids
aqueous solutions
Water
Carboxylic acids
induction
methodology
water
primary amines
dosage
avidin
Polylysine
Avidin

ASJC Scopus subject areas

  • Radiological and Ultrasound Technology
  • Radiology Nuclear Medicine and imaging

Cite this

A method for detection of hydroxyl radicals in the vicinity of biomolecules using radiation-induced fluorescence of coumarin. / Makrigiorgos, G. M.; Baranowska-Kortylewicz, J.; Bump, E.; Sahu, S. K.; Berman, R. M.; Kassis, A. I.

In: International Journal of Radiation Biology, Vol. 63, No. 4, 01.01.1993, p. 445-458.

Research output: Contribution to journalArticle

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