An isotopic assay of dUTPase activity based on coupling with thymidylate synthase.

B Gołos; W Rode

doi:10.18388/abp.1999_4155

An isotopic assay of dUTPase activity based on coupling with thymidylate synthase.

B Gołos Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warszawa.;
W Rode

DOI: https://doi.org/10.18388/abp.1999_4155

Abstract

A new rapid, sensitive and convenient procedure is presented allowing determination of dUTPase activity. With [5-(3)H]dUTP used as the substrate, dUTPase, converts it to the corresponding monophosphate and is coupled with thymidylate synthase-catalyzed reaction, resulting in tritium release from [5-(3)H]dUMP. Following charcoal absorption of the labeled nuleotides, radioactivity of tritiated water is determined. The new assay was tested to show comparable results with a previously described assay, based on measuring dUTPase-catalyzed [5-(3)H]dUMP production.

Published

1999-09-30

Issue

Vol. 46 No. 3 (1999)

Section

Articles

Acta Biochimica Polonica is an OpenAccess quarterly and publishes four issues a year. All contents are distributed under the Creative Commons Attribution-ShareAlike 4.0 International (CC BY 4.0) license. Everybody may use the content following terms: Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.