Isolation of a Ca-dependent erythrolytic protein (perforin) from cytotoxic T-lymphocytes.
A Ca-dependent erythrolytic protein (perforin) was isolated from a cytotoxic T-cell line (CTLL2). Cellular extracts were fractionated on DEAE-cellulose and hydrophobic Phenyl-Sepharose columns. Lytic activity was tightly bound to the hydrophobic column and was eluted with 50% ethyleneglycol. The erythrolytic activity was dependent on the concentration of Ca2+ ions, and heparin accelerated the lysis of erythrocytes by perforin 10-fold, with a half maximal concentration of 12 ng/ml. The activity was strongly inhibited by micromolar concentrations of heavy metal ions, such as Zn2+ and Fe2+, and glycylarginine-methylcoumarinamide (Gly-Arg-MCA) in the presence of 100 ng/ml heparin.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D006461:Hemolysis; D007700:Kinetics; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D051379:Mice; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D012756:Sheep; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D052899:Pore Forming Cytotoxic Proteins; D054353:Perforin; D002118:Calcium