title: Purification and characterization of activated human erythrocyte prolidase.
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.
authors.affiliation: MRC Human Genetics Group, Montreal Children's Hospital, Qué., Canada.
authors.forename: A M
authors.initials: AM
authors.lastname: Richter
authors.forename: G L
authors.initials: GL
authors.lastname: Lancaster
authors.forename: F Y
authors.initials: FY
authors.lastname: Choy
authors.forename: P
authors.initials: P
authors.lastname: Hechtman
mesh_terms: D001798:Blood Proteins / Q000032:analysis; D015153:Blotting, Western; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004150:Dipeptidases / Q000097:blood* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D015151:Immunoblotting; D008345:Manganese / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D001798:Blood Proteins; D008345:Manganese; D004150:Dipeptidases; C022449:proline dipeptidase