Partial purification and properties of the two common inherited forms of human erythrocyte adenylate kinase

• DOI: 10.1042/bj1300797
• Publication Date: 2015-08-10T19:47:58Z
• Authors: C. Brownson; N. Spencer
• Keywords: Chromatography; Protein Denaturation; 0303 health sciences; Erythrocytes; Hot Temperature; Electrophoresis, Starch Gel; Phosphotransferases; Genetic Variation; Iodoacetates; Buffers; Hydrogen-Ion Concentration; Blood Protein Electrophoresis; Amides; Adenosine Diphosphate; Isoenzymes; Molecular Weight; 03 medical and health sciences; Humans; Magnesium

1. The partial purification of adenylate kinase, types 1 and 2, from human erythrocytes is described. 2. Gel chromatography of both forms of the enzyme gave estimates of the molecular weights in the range 20000–23000. 3. Studies on crude haemolysates at various pH values indicated that the type 2 enzyme was less stable than the type 1. Heat denaturation studies on the partially purified enzymes confirmed these findings. 4. Measurements of rates of inhibition by iodoacetate and iodoacetamide showed that the type 2 enzyme reacts more readily than the type 1 enzyme with both reagents. 5. The effect of temperature on the initial velocity of ADP formation was measured at a single concentration of both AMP and MgATP2-. The two forms of the enzyme responded differently to increasing temperature.