Pyrimidine Nucleoside Monophosphate Kinase from Rat Liver and Rat Novikoff Ascites Hepatoma (EC 2.7.4.14)

Antonio Orengo, Patricia Maness

Research output: Research - peer-reviewArticle

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Abstract

This chapter describes the pyrimidine nucleoside monophosphate kinase from rat liver and rat Novikoff ascites hepatoma. The enzyme occupies a strategic position in the biosynthesis of pyrimidine nucleotides because its phosphate-acceptor substrates are products of both the de novo and the salvage pathways. The phosphate-acceptor specificity is peculiar because the enzymes catalyzes the phosphorylation of cytidine monophosphate (CMP), deoxycytidine monophosphate (dCMP), and uridine monophosphate (UMP) but not deoxyuridine monophosphate (dUMP) that differs from UMP by only one atom of oxygen just as dCMP differs from CMP. Two methods of assay are used. The radiochemical assay measures the conversion of nucleoside monophosphate to nucleoside diphosphate or the transfer of γ-phosphate from adenosine tri phosphate (ATP) to the nucleoside monophosphate by separation of reactants and products on paper electrophoresis. In the latter assay, pyruvate kinase is coupled with lactate dehydrogenase to measure adenosine diphosphate (ADP) formation. When ATP or deoxyadenosine triphosphate (dATP) serve as phosphate donor, substrate inhibition occurs at concentrations of CMP greater than 0.13 mM.

LanguageEnglish (US)
Pages321-331
Number of pages11
JournalMethods in Enzymology
Volume51
Issue numberC
DOIs
StatePublished - Jan 1 1978

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Experimental Liver Neoplasms
Ascites
Phosphates
Liver
cytidylate kinase
Rats
Cytidine Monophosphate
Nucleosides
Assays
Deoxycytidine Monophosphate
Uridine Monophosphate
Adenine Nucleotides
Enzymes
Substrates
Pyrimidine Nucleotides
Paper Electrophoresis
Deoxyuridine
Pyruvate Kinase
Diphosphates
L-Lactate Dehydrogenase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Pyrimidine Nucleoside Monophosphate Kinase from Rat Liver and Rat Novikoff Ascites Hepatoma (EC 2.7.4.14). / Orengo, Antonio; Maness, Patricia.

In: Methods in Enzymology, Vol. 51, No. C, 01.01.1978, p. 321-331.

Research output: Research - peer-reviewArticle

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