Purification, characterization, and localization of an ATP diphosphohydrolase in porcine kidney

Raf Lemmens, Luc Kupers, Jean Sévigny, Adrien R. Beaudoin, Gilles Grondin, Agnes Kittel, Etienne Waelkens, Luc Vanduffel

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Membranes of pig kidney cortex tissue were solubilized in the presence of Triton X-100. Partial purification of ATP diphosphohydrolase (ATPDase) was achieved by successive chromatography on concanavalin A-Sepharose, Q- Sepharose Fast Flow, and 5'-AMP-Sepharose 4B. Monoclonal antibodies against ATPDase were generated. Further purification of the ATPDase was obtained by immunoaffinity chromatography with these monoclonal antibodies. NH2-terminal amino acid sequencing of the 78-kDa protein showed a sequence very homologous to mammalian CD39. The protein is highly glycosylated, with a nominal molecular mass of ~57 kDa. The purified enzyme hydrolyzed di- and triphosphates of adenosine, guanosine, cytidine, uridine, inosine, and thymidine, but AMP and diadenosine polyphosphates could not serve as substrates. All enzyme activities were dependent on divalent cations and were partially inhibited by 10 mM sodium azide. The distribution of the enzyme in pig kidney cortex was examined immunohistochemically. The enzyme was found to be present in blood vessel walls of glomerular and peritubular capillaries.

Original languageEnglish
Pages (from-to)F978-F988
JournalAmerican Journal of Physiology - Renal Physiology
Issue number6 47-6
Publication statusPublished - Jun 1 2000



  • Apyrase
  • CD39
  • Nucleoside triphosphate diphosphohydrolase
  • Triton X-100

ASJC Scopus subject areas

  • Physiology
  • Urology

Cite this

Lemmens, R., Kupers, L., Sévigny, J., Beaudoin, A. R., Grondin, G., Kittel, A., Waelkens, E., & Vanduffel, L. (2000). Purification, characterization, and localization of an ATP diphosphohydrolase in porcine kidney. American Journal of Physiology - Renal Physiology, 278(6 47-6), F978-F988.