Adenosine deaminase isoenzymes of the opossum Didelphis virginiana: initial chromatographic and kinetic studies

John G. Niedzwicki, Caspar Liou, Darrell R. Abernethy, Joseph E. Lima, Anne Hoyt, Michael Lieberman, Nicholas C. Bethlenfalvay

Research output: Contribution to journalArticlepeer-review

Abstract

Extracts of liver and spleen were used to isolate opossum adenosine deaminase isoenzymes (ADA, and ADA2) and to determine their activities with adenosine and 2′-deoxyadenosine as substrates. Km values (μM) for adenosine and 2′-deoxyadenosine, respectively, as substrates for partially purified opossum liver adenosine deaminase isoenzymes were ADA,: 57 ± 7 vs. 26 ± 4 and ADA2: 285 ± 25 vs. 580 ± 92. In crude spleen extract, ADA2 activity was stable at 56°C during 40 min of incübation. ADA1 activity declined in a linear fashion under the above conditions with an apparent T 1 2 of 80 min. Sephadex G-150 column chromatography of crude spleen extract showed the apparent molecular weight of the ADA activity not inhibited by (±)-EHNA (i.e. ADA2) to be 170 kDa; ADA activity fully inhibited by (±)-EHNA (i.e. ADA,) eluted in the fractions corresponding to a molecular weight of 35 kDa.

Original languageEnglish (US)
Pages (from-to)291-298
Number of pages8
JournalComparative Biochemistry and Physiology -- Part B: Biochemistry and
Volume111
Issue number2
DOIs
StatePublished - Jun 1995

Keywords

  • ADA1
  • ADA2
  • Adenosine deaminase
  • EHNA
  • Isoenzymes
  • Opossum
  • Severe combined immunodeficiency disease

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology

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