Purification and characterization of activated human erythrocyte prolidase |
| |
| Authors: | A M Richter G L Lancaster F Y Choy P Hechtman |
| |
| Affiliation: | MRC Human Genetics Group, Montreal Children's Hospital, Qué., Canada. |
| |
| Abstract: | Prolidase (E.C. 3.4.13.9) has been purified 7500-fold to homogeneity from human erythrocytes in a Mn2+-activated form using conventional and fast protein liquid chromatography columns. The procedure includes a 1-h incubation of the crude hemolysate at 50 degrees C with 1 mM MnCl2. Following this novel step, prolidase retains full activity, obviating the requirement for preincubation of each enzyme fraction with Mn2+ prior to assay. Preincubation with MnCl2 does not change the isoelectric point of the enzyme. The molecular weight of the purified enzyme was 58,000 when measured by SDS-PAGE. Western blotting, using rabbit antibody raised to human kidney prolidase, with partially purified erythrocyte enzyme revealed a cross-reacting band at Mr 58,000. |
| |
| Keywords: | |
|
|
