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Purification of Myocardial Adenosine Kinase Using Affinity and Ion-Exchange Chromatography

  • Martin P. Uitendaal
  • Jan W. De Jong
  • Eef Harmsen
  • Elisabeth Keijzer
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 122B)

Abstract

Myocardial adenosine kinase (AK; EC 2.7.1.20) presumably plays a key role in the maintenance of adequate adenine nucleotide levels in the heart cell1–3. In order to study this enzyme in detail, we purified rat-heart AK to apparent homogeneity after a previous report on partial purification from this source4. The method presented here includes elution of AK from a 5′-AMP-Sepharose 4B column with a buffer containing adenosine. The endogenous adenosine in the fractions altered the specific activity of the radioactive substrate in the AK assay. This could be corrected for by means of HPLC adenosine measurements.

Keywords

Adenosine Deaminase Cytoplasmic Fraction Purification Factor Sepharose Column Endogenous Adenosine 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • Martin P. Uitendaal
    • 1
  • Jan W. De Jong
    • 1
  • Eef Harmsen
    • 1
  • Elisabeth Keijzer
    • 1
  1. 1.Cardiochemical Laboratory, ThoraxcenterErasmus University RotterdamRotterdamThe Netherlands

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