PMID: 6414331Jul 15, 1983Paper

A 13C tracer method for quantitating de novo pyrimidine biosynthesis in vitro and in vivo

Analytical Biochemistry
J M StrongR L Cysyk

Abstract

Gas chromatographic/mass spectrometric methods for the measurement of the flux through the de novo pyrimidine biosynthetic pathway by quantitating the incorporation of [13C]bicarbonate and 13CO2 into the uracil nucleotide pool in L1210 tumors are reported. Simultaneous measurements of the incorporation of [13C]bicarbonate and the more commonly used [14C]bicarbonate into uridine of L1210 cells in vitro showed that the two methods were comparable. A modification of the method was applied to in vivo studies where the incorporation of 13CO2 into the uracil nucleotide pool of L1210 tumors in mice was quantitated. The measurements were used to determine changes in the flux through the de novo pyrimidine pathway in animals pretreated with known inhibitors of the pathway. A comparison of control animals with those pretreated with 6-azauridine, acivicin, and pyrazofurin resulted in mean percentage inhibitions of 87, 95, and 94%, respectively. This technique should allow investigation of the respective contributions of salvage and de novo synthesis in the formation of pyrimidines in vivo and the effects of agents designed as enzyme inhibitors of the de novo pathway.

References

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Citations

Apr 30, 1990·European Journal of Biochemistry·D W ZaharevitzR L Cysyk
May 5, 2018·Cell·Cholsoon JangJoshua D Rabinowitz
May 26, 1998·Analytical Chemistry·P Chen, F P Abramson

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