Serologic assay of DNA base damage. I. 5-hydroxymethyldeoxyuridine, a radiation product of thymidine. [Gamma radiation]

Journal Article · Tue Aug 01 04:00:00 EDT 1978 · Radiat. Res.; (United States)

DOI:https://doi.org/10.2307/3574906· OSTI ID:6204025

Lewis, H L; Muhleman, D R; Ward, J F

We have developed a serologic assay for 5-hydroxymethyldeoxyuridine, (5-HMUdR) an ionizing radiation product of thymidine. Antibodies were prepared to a bovine serum albumin conjugate of 5-hydroxymethyluridine (5-HMUR). Using phage neutralization assay 0.1 nmole of 5-HMUR can be detected. The antiserum cross-reacted with thymidine but with no other naturally occurring DNA components. There was cross-reaction with uridine, 6-methyluracil, and the halogenated analogs of thymidine. Interference by cross-reaction with thymidine was eliminated by saturation of the assay with excess thymidine. Three micromolar (0.3 nmole/0.1 ml) 5-HMUR can be detected in the presence of 8 mM thymidine. The assay was applied to the estimation of 5-HMUdR produced by ..gamma.. irradiation of thymidine in O/sub 2/. The G value for product formation was 0.05. Irradiation at pH 14 greatly increased the yield. There was a threefold increase in G value when irradiated samples were heated at pH 9 or treated with 10/sup -3/ M ferrous ions, postirradiation procedures which convert thymidine methylperoxides to 5-HMUdR. This suggests that cross-reaction with the peroxide is not extensive. Estimation of 5-HMUdR in DNA irradiated in O/sub 2/ gave a G value roughly the same as that found in irradiation of thymidine. This yield is based on the assumption that antibody reacts equally with the 5-HMU moiety in DNA and as the mononucleoside. 5-HMUdR was detected as one nucleoside per 3 x 10/sup 6/ daltons of DNA.

• Research Organization: Univ. of California, Los Angeles
• OSTI ID: 6204025
• Journal Information: Radiat. Res.; (United States), Journal Name: Radiat. Res.; (United States) Vol. 75:2; ISSN RAREA
• Country of Publication: United States
• Language: English

Related Subjects

550200 -- Biochemistry
560111* -- Radiation Effects on Biochemicals-- In Vitro-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANTIBODIES
AZINES
BIOASSAY
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
DEOXYURIDINE
DNA
ELECTROMAGNETIC RADIATION
GAMMA RADIATION
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
IMMUNE SERUMS
IONIZING RADIATIONS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PYRIMIDINES
RADIATION EFFECTS
RADIATIONS
RIBOSIDES
THYMIDINE
URACILS