Purification of human interleukin 2 to apparent homogeneity and partial characterization of its receptor.
Human interleukin 2, produced by phytohemagglutinin-stimulated peripheral blood lymphocytes cultured in the absence of serum, has been purified to apparent homogeneity with a two-step combination of affinity chromatography with Cibacron blue-Sepharose and gel filtration. The specific activity of the purified IL-2 was 620,000 U/mg of protein, with a total recovery of approximately 13% biological activity. Purified IL-2 stimulated lymphocyte proliferation at a concentration of 4.3 X 10(-10) M. IL-2 was radiolabeled with reductive methylation to a specific activity of 350 microCi/nmol. Binding studies with phytohemagglutinin-activated lymphocytes, known to express receptor for IL-2, identified a single population of approximately 21,000 receptors per cell with an equilibrium dissociation constant of 7.1 X 10(-10) M.
D001345:Autoradiography; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007376:Interleukin-2 / Q000302:isolation & purification*; D007768:Lactalbumin / Q000494:pharmacology; D010835:Phytohemagglutinins / Q000494:pharmacology; D011971:Receptors, Immunologic / Q000302:isolation & purification*; D015375:Receptors, Interleukin-2; D012685:Sepharose / Q000031:analogs & derivatives; D013601:T-Lymphocytes / Q000378:metabolism
D007376:Interleukin-2; D010835:Phytohemagglutinins; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; C017842:cibacron blue-sepharose; D012685:Sepharose; D007768:Lactalbumin; D004229:Dithiothreitol