title: Human leukocyte glucose-phosphate-isomerase purification by affinity elution and immunological study.
abstract: The authors have purified glucose phosphate isomerase (GPI) from human leukocytes ; they used as starting material leukemic leukocytes obtained from a patient with hyper-leukocytic acute myeloid leukemia ; it was possible to obtain several milligrams of pure enzyme from a single patient. The purification procedure includes a two step precipitation by ammonium sulfate and one column chromatography on a cation exchanger with specific elution by 6 phosphogluconate, a ligand of GPI ; of the two cation exchangers tested, phosphocellulose was found to lead to a better yield than CM-Sephadex. The end product of purification had a specific activity of 855 UI/mg and gave only one band in sodium dodecyl sulphate polyacrylamide gel electrophoresis. Anti GPI monospecific rabbit serum was obtained with purified enzyme. GPI from the various blood cells of ten normal controls was studied immunologically and the ratio of the enzymatic activity to the immunological reactivity was measured ; this ratio (i.e. the molecular specific activity) was lower in granulocytes than in lymphocytes and still more depressed in platelets and hemolysate. The significance of such differences is discussed.
mesh_terms: D001792:Blood Platelets / Q000201:enzymology; D002846:Chromatography, Affinity; D004912:Erythrocytes / Q000201:enzymology; D005956:Glucose-6-Phosphate Isomerase / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification; D006098:Granulocytes / Q000201:enzymology; D006801:Humans; D005779:Immunodiffusion; D007525:Isoelectric Focusing; D015470:Leukemia, Myeloid, Acute / Q000201:enzymology; D007962:Leukocytes / Q000201:enzymology*
chemical_list: D005956:Glucose-6-Phosphate Isomerase