Thin-layer chromatography of thymidine and thymidine metabolites in enzyme assays and cell extracts

Bols, Niels C.; Bowen, Bill; Khor, Kirin; Boliska, S.A.

doi:10.1016/0003-2697(80)90142-6

Cited by 12 publications

(3 citation statements)

References 17 publications

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“…It was obvious from the present data that there is an efficient catabolism of 13H]TdR. This observation is in accordance with several other reports of high amounts of acid-soluble radioactivity following injection of 13HITdR (Cronkite et al, 1962;Hinrichs, Petersen & Baserga, 1964: Tew & Taylor, 1978Carlsson, Johanson & Safwenberg, 1979;Bols et al, 1980). The great acid-soluble portion of radioactivity consists of tritiated water, labelled TdR catabolites not available for DNA synthesis, unchanged t3H1TdR and labelled metabolites still available for DNA synthesis.…”

Section: Discussionsupporting

confidence: 93%

“…It is well known that only a minor fraction of the exogenously administered TdR will be used for incorporation into DNA (Hopkins & Wakefield, 1977;Tew & Taylor, 1978). The main degradation products of TdR are known to be P-aminoisobutyric acid and P-ureidoisobutyric acid, which are further metabolized to CO,, NH,, and water (Tew & Taylor, 1978;Bols et al, 1980). Tritiated water is therefore one of the major labelled products seen after administration of [6-)HITdR.…”

Section: B Hellman and S Ullbergmentioning

confidence: 99%

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Rate of Incorporation of R3hlthymidine In Various Tissues of the Mouse A basis for the evaluation of genotoxic effects of chemicals

Hellman

Ullberg

1986

Cell Proliferation

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13HlThymidine has been extensively used as a selective pr6cursor to DNA in studies on the kinetics of cell proliferation. We have become interested in measuring early inhibition of the DNA synthesis in various organs of intact animals for detecting genotoxic properties of chemicals. Such experiments should, for convenience and to achieve a large capacity, be performed in the simplest way possible.The present paper deals with some practical aspects on the use of I3H]thymidine in uivo. C6-3HlThymidine was injected intraperitoneally in mice and the uptake of

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Section: Discussionsupporting

confidence: 93%

Section: B Hellman and S Ullbergmentioning

confidence: 99%

Rate of Incorporation of R3hlthymidine In Various Tissues of the Mouse A basis for the evaluation of genotoxic effects of chemicals

Hellman

Ullberg

1986

Cell Proliferation

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“…Previously the separation of all four compounds in the catabolism of uracil required two-dimensional chromatography (Fink et al, 1956). One-dimensional procedures failed to resolve uracil from dihydrouracil (Fritzson, 1962;Ikenaka et al, 1979) or NC/3A from /3-alanine (Fritzon, 1962;Sanno et al, 1970;Bols et al, 1980). A recent report describes the complete separation of the four fluoro analogues of these uracil metabolites by high-pressure liquid chromatography (Sommadossi et al, 1982).…”

Section: Discussionmentioning

confidence: 99%

Pyrimidine catabolism: individual characterization of the three sequential enzymes with a new assay

Traut

Loechel²

1984

Biochemistry

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We have developed a one-dimensional thin-layer chromatography procedure that resolves the initial substrate uracil and its catabolic derivatives dihydrouracil, N-carbamoyl-beta-alanine (NCBA) and beta-alanine. This separation scheme also simplifies the preparation of the radioisotopes of N-carbamoyl-beta-alanine and dihydrouracil. Combined, these methods make it possible to assay easily and unambiguously, jointly or individually, all three enzyme activities of uracil catabolism: dihydropyrimidine dehydrogenase, dihydropyrimidinase, and N-carbamoyl-beta-alanine amidohydrolase. Earlier reports had presented data suggesting that these three enzyme activities were combined in a complex because they appeared to be controlled at a single genetic locus [Dagg, C. P., Coleman, D.L., & Fraser, G.M. (1964) Genetics 49, 979-989] and because they appeared able to channel metabolites [Barrett, H.W., Munavalli, S.N., & Newmark, P. (1964) Biochim. Biophys. Acta 91, 199-204]. Although the three enzymes from rat liver have similar sizes, with apparent molecular weights of 218 000 for dihydropyrimidine dehydrogenase, 226 000 for dihydropyrimidinase, and 234 000 for NC beta A amidohydrolase, they are easily separated from each other. Kinetic studies show no evidence of substrate channeling and therefore do not support a model for an enzyme complex. The earlier reports may be explained by our studies on the amidohydrolase, which suggest that under certain conditions this enzyme may become the rate-limiting step in uracil catabolism.

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Regulation of Extracellular Thymidine Pools in Human Blood Samples

Pero

Olsson

Johnsonf

1985

Genetic Consequences of Nucleotide Pool Imbalance

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Thin-layer chromatography of thymidine and thymidine metabolites in enzyme assays and cell extracts

Cited by 12 publications

References 17 publications

Rate of Incorporation of R3hlthymidine In Various Tissues of the Mouse A basis for the evaluation of genotoxic effects of chemicals

Rate of Incorporation of R3hlthymidine In Various Tissues of the Mouse A basis for the evaluation of genotoxic effects of chemicals

Pyrimidine catabolism: individual characterization of the three sequential enzymes with a new assay

Regulation of Extracellular Thymidine Pools in Human Blood Samples

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