title: Immunochemical studies of human prolidase with monoclonal and polyclonal antibodies: absence of the subunit of prolidase in erythrocytes from a patient with prolidase deficiency.
abstract: Prolidase was highly purified from human liver and erythrocytes. NaDodSO4/acrylamide gel electrophoresis revealed that these preparations contained a major protein with MW = 56,000. The mass of prolidase was estimated on gel filtration to be MW = 97,000, for both enzyme preparations. A monoclonal antibody was raised against the liver enzyme and a specific antiserum against the erythrocyte enzyme. The monoclonal antibody (EP-2) recognized prolidase from erythrocytes and liver, in equal proportions. The antiserum also recognized the enzyme from erythrocytes and liver. Immunoprecipitation studies with these antibodies suggested only a single species of prolidase in erythrocytes and liver. Using an immobilized monoclonal antibody (EP-2) as an immunoadsorbent, prolidase was partially purified from crude extracts, and the protein of the partially purified enzyme was identified by immunoblotting using antiserum. A protein band with a MW = 56,000 was demonstrated specifically when crude extracts from the liver and erythrocytes were examined using NaDodSO4/acrylamide gel electrophoresis. The subunit protein was absent in erythrocytes from a patient with prolidase deficiency. We propose that the absence of the subunit is one cause of the prolidase deficiency.
authors.affiliation: Department of Pediatrics, Kumamoto University Medical School, Japan.
authors.forename: F
authors.initials: F
authors.lastname: Endo
authors.forename: K
authors.initials: K
authors.lastname: Motohara
authors.forename: Y
authors.initials: Y
authors.lastname: Indo
authors.forename: I
authors.initials: I
authors.lastname: Matsuda
mesh_terms: D000911:Antibodies, Monoclonal; D004150:Dipeptidases / Q000172:deficiency / Q000276:immunology* / Q000302:isolation & purification; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D007120:Immunochemistry; D007163:Immunosorbent Techniques; D008099:Liver / Q000201:enzymology; D008970:Molecular Weight; D011487:Protein Conformation
chemical_list: D000911:Antibodies, Monoclonal; D004150:Dipeptidases; C022449:proline dipeptidase