5′-Nucleotidases: Specific assays for five different enzymes in cell extracts

Chiara Rampazzo, Cristina Mazzon, Peter Reichard, Vera Bianchi

Research output: Contribution to journalArticle

Abstract

Several mammalian 5′-nucleotidases (5-NTs), attached to membranes or present in the cytosol or in mitochondria, remove the phosphate from ribo- and deoxyribonucleotides with different specificities for the sugar and base moieties. Some enzymes probably participate in signaling functions by producing adenosine from AMP. A more general function may be to prevent overproduction of deoxyribonucleotides. 5-NTs may affect the pharmacological activity of nucleoside analogs and also be involved in their mitochondrial toxicity. Here we describe for five cloned 5-NT specific assays that largely rely on new inhibitors for some of the enzymes. The assays can be used to quantitate each enzyme in crude cell extracts. To ascertain their validity we applied each assay to extracts from genetically modified cells that overproduce separately each of the five enzymes. The methodology should be useful in further studies of the physiological function of 5-NTs and their influence on the clinical use of nucleoside analogs.

Original languageEnglish
Pages (from-to)258-263
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume293
Issue number1
DOIs
Publication statusPublished - 2002

Keywords

  • Enzyme overproduction
  • Nucleoside analogs
  • Signaling
  • Substrate cycles

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Fingerprint Dive into the research topics of '5′-Nucleotidases: Specific assays for five different enzymes in cell extracts'. Together they form a unique fingerprint.

  • Cite this