Orotate phosphoribosyltransferase: Orotidylate decarboxylase (Ehrlich Ascites Cell)

Mary Ellen Jones, Prabhakar Rao Kavipurapu, Thomas W. Traut

Research output: Research - peer-reviewArticle

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Abstract

This chapter describes purification procedure related to the orotate phosphoribosyltransferase enzyme. Assay procedures reported include methods measuring the spectrophotometric changes that occur when a pyrimidine base is converted to a nucleotide or when orotidylic acid (OMP) is converted to uridylic acid (UMP), methods that separate the free base from the nucleotides, and methods measuring the release of [14C]O2 from [14C]carboxyl-labeled orotate or orotidylate. This assay is appropriate for both crude or pure enzyme preparations. In this procedure orotate, OMP, and UMP are separated from one another by thin-layer chromatography. It is also useful when one wishes to estimate how effective inhibitors of the decarboxylase are when the OMP level is that maintained at a steady-state level by the complex itself,for example, when orotate and phosphoribosyl pyrophosphate (P-Rib-PP) are the substrates and only the OMP formed by the transferase is present, this assay is ideal.

LanguageEnglish (US)
Pages155-167
Number of pages13
JournalMethods in Enzymology
Volume51
Issue numberC
DOIs
StatePublished - Jan 1 1978

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Orotidine-5'-Phosphate Decarboxylase
Orotate Phosphoribosyltransferase
Uridine Monophosphate
Assays
Ascites
Nucleotides
Enzymes
Phosphoribosyl Pyrophosphate
Thin layer chromatography
Transferases
Purification
Substrates
pyrimidine
orotidylic acid
Thin Layer Chromatography

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Orotate phosphoribosyltransferase : Orotidylate decarboxylase (Ehrlich Ascites Cell). / Jones, Mary Ellen; Kavipurapu, Prabhakar Rao; Traut, Thomas W.

In: Methods in Enzymology, Vol. 51, No. C, 01.01.1978, p. 155-167.

Research output: Research - peer-reviewArticle

Jones, Mary Ellen ; Kavipurapu, Prabhakar Rao ; Traut, Thomas W./ Orotate phosphoribosyltransferase : Orotidylate decarboxylase (Ehrlich Ascites Cell). In: Methods in Enzymology. 1978 ; Vol. 51, No. C. pp. 155-167
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