title: Purification of L-type pyruvate kinase from human liver by affinity chromatography on Blue-Dextran-Sepharose column.
abstract: L-type pyruvate kinase (ATP: pyruvate 2-O-phosphotransferase; EC 2.7.1.40) was purified from human liver by an original method. This purification included toluene extraction, a-monium sulphate fractionation, DEAE-Sephadex bactchwise, CM-Sephadex batchwise with elective elution by ATP and affinity chromatography on a Blud Dextran-Sepharose column with specific elution by fructose 1, 6-diphosphate. This purification procedure allowed us to obtain 6 mg protein with a specific activity of 420 IU/mg protein, i.e. 2,690-fold purification with an overall yield of 34%. This preparation was homogeneous as judged by immuno-diffusion, acrylamide and sodium dodecyl sulphate acrylamide-gel electrophoresis.
mesh_terms: D002846:Chromatography, Affinity; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D011770:Pyruvate Kinase / Q000302:isolation & purification*
chemical_list: D007527:Isoenzymes; D011770:Pyruvate Kinase