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1999 | 46 | 3 |
Tytuł artykułu

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

Autorzy
Warianty tytułu
Języki publikacji
EN
Abstrakty
EN
A new rapid, sensitive and convenient procedure is presented allowing determination of dUTPase activity. With [5-3H]dUTP used as the substrate, dUTPase, converts it to the corresponding monophosphate and is coupled with a thymidylate synthase-catalyzed reaction, resulting in tritium release from [5-3H]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 [5-3H]dUMP production.
Wydawca
-
Rocznik
Tom
46
Numer
3
Opis fizyczny
p.837-840,fig.
Twórcy
autor
  • Polish Academy of Sciences, L.Pasteura 3, 02-093 Warsaw, Poland
autor
Bibliografia
  • 1. Vilpo, J.A. & Autio-Harmainen, H. (1983) Ura- cil-DNA glycosylase and deoxyuridine triphos­phatase: Studies of activity and subcellular lo­cation in human normal and malignant lym­phocytes. Scand. J. Clin. Lab. Invest. 43, 583- 590.
  • 2. Carreras, C.W. & Santi, D. (1995) The cata­lytic mechanism and structure of thymidylate synthase. Annu. Rev. Biochem. 64, 721-762.
  • 3. Hokari, S., Sakagiahi, Y. & Tukada, K. (1982) Enhanced activity of deoxyuridine 5'-triphos­phatase in regenerating rat liver. Biochem. Biophys. Res. Commun. 108, 95-101.
  • 4. Shlomai, M.A. & Romberg, A. (1978) Deoxy­uridine triphosphatase of Escherichia coli. J. Biol Chem. 253, 3305-3312.
  • 5. Williams, M.V. & Cheng, Y-c. (1979) Human deoxyuridine triphosphate nucleotidohy- drola8e. Purification and characterization of the deoxyuridine triphosphate nucleotidohy- drolase from acute lymphocytic leukemia. J. Biol Chem. 254, 2897-2901.
  • 6. Greenberg, G.R. & Somerville, R.L. (1962) Deoxyuridylate kinase and deoxyuridinetri- phosphatase in Escherichia coli. Biochemistry 48, 247-257.
  • 7. Roberts, D.W. (1966) An isotopic assay for thymidylate synthetase. Biochemistry 5,3546- 3548.
  • 8. Higgins, G.M. & Anderson, R.M. (1931) Exper­imental pathology of the liver. Arch. Pathol. 12, 186-202.
  • 9. Ciesla, J., Golos, B., Dzik, J.M., Pawelczak, K.,. Kempny, M., Makowski, M., Bretner, M., Kulikowski, T., Machnicka, B., Rzeszotarska, B. & Rode, W. (1995) Thymidylate synthases from Hymenolepis diminuta and regenerating rat liver: Purification, properties, and inhibi­tion by substrate and cofactor analogues. Biochim. Biophys. Acta 1249, 127-136.
  • 10. Rode, W., Scanlon, K.J., Hynes, J. & Bertino, J.R. (1979) Purification of mammalian tumor (L1210) thymidylate synthetase by affinity chromatography on stable biospecific adsor­bent. Stabilization of the enzyme with neutral detergents. J. Biol Chem. 254,11538-11543.
  • 11. Spector, T. (1978) Refinement of the Coomas- sie Blue method of protein quantitation. Anal. Biochem. 86, 142-146.
Typ dokumentu
Bibliografia
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bwmeta1.element.agro-article-10a7c398-005b-4fac-bf9f-59ec84dc28f6
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