Ishiura S, Anraku H, Kamo I, Koizumi H, Arahata K, Sugita H
National Institute of Neuroscience, NCNP, Tokyo.
J Biochem. 1987 Jul;102(1):9-12. doi: 10.1093/oxfordjournals.jbchem.a122045.
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.
从细胞毒性T细胞系（CTLL2）中分离出一种钙依赖性红细胞溶解蛋白（穿孔素）。细胞提取物在DEAE - 纤维素柱和疏水苯基 - 琼脂糖柱上进行分级分离。溶血活性紧密结合在疏水柱上，并用50%乙二醇洗脱。红细胞溶解活性依赖于Ca2+离子的浓度，肝素可使穿孔素介导的红细胞溶解加速10倍，半数最大浓度为12 ng/ml。在100 ng/ml肝素存在的情况下，微摩尔浓度的重金属离子，如Zn2+和Fe2+，以及甘氨酰精氨酸 - 甲基香豆素酰胺（Gly - Arg - MCA）可强烈抑制该活性。
