Purification of Myocardial Adenosine Kinase Using Affinity and Ion-Exchange Chromatography
Myocardial adenosine kinase (AK; EC 220.127.116.11) 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.
KeywordsAdenosine Deaminase Cytoplasmic Fraction Purification Factor Sepharose Column Endogenous Adenosine
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