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Nucleic Acids Research, 1978, Vol. 5, No. 12 4753-4760
© 1978

Articles

2'-Deoxy-2'-fluorouridine-5'-phosphate: an alternative substrate for thymidylate synthetase from Escherichia coli K12

Franz Wohlrab, Tomasz Haertlé$, Thomas Trichtinger and Wilhelm Guschlbauer

Service de Biochimie, Bat. 142, Departement de Biologie, Centre d'Etudes Nucléaires de Saclay B.P. No.2, F-91190 Gif-sur-Yvette, France

Received November 3, 1978.

The substrate specificity of 2'-deoxy-2'-substituted uridines and their 5'-phosphates towards thymidylate synthetase from Escherichia coli K12 was investigated. Besides the natural substrate 2'-deoxyuridine-5'-phosphate (dUMP), only 2'-deoxy-2'-fluorouridine-5'-phosphate (dUfIMP) was a substrate. The KM of dUfIMP is 11 times higher than that of dUMP, while the Vmax values are virtually the same. It is concluded that the size of the 2'-substituent and not its polarity (and the concomitant conformational change) determines substrate specificity of thymidylate synthetase.

$Present address: Institute of Biochemistry, University Adam Mickiewicz, Poznan Poland


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