Purification to apparent homogeneity of murine interleukin 1.
Purification to apparent homogeneity of murine interleukin 1.
复制标题
纯化小鼠白细胞介素 1，使其表现出同质性。
作者：
S. Mizel;D. Mizel
从P388 D1细胞系巨噬细胞的培养液中，通过硫酸铵分级分离、苯基琼脂糖凝胶和Ultrogel AcA 54层析以及制备性等电聚焦，将白细胞介素1纯化至表观均一性。白细胞介素1纯化超过300倍，比活性约为1 × 10（6）U/mg蛋白。纯化的白细胞介素1具有微异质性。三种电荷物种，我们称之为α，β，γ，解决了三甘氨酸不连续聚丙烯酰胺凝胶电泳。测定白细胞介素1 α类的部分氨基酸含量。根据纯化数据，似乎白细胞介素1在10（-11）至10（-10）M浓度范围内具有生物活性。纯化的成功部分是由于使用了超诱导方案，该方案导致白细胞介素1以约1微克/24小时/4 × 10（6）P388 D1细胞的速率产生。
Interleukin 1 was purified to apparent homogeneity from the culture fluid of P388D1 cell line macrophages by a procedure involving ammonium sulfate fractionation, phenyl Sepharose and Ultrogel AcA54 chromatographies, and preparative isoelectrofocusing. The interleukin 1 was purified more than 300-fold to a specific activity of approximately 1 X 10(6) U/mg protein. The purified interleukin 1 exhibited microheterogeneity. Three charge species, which we have termed alpha, beta, gamma, were resolved by Tris-glycinate discontinuous polyacrylamide gel electrophoresis. The partial amino acid content of the alpha species of interleukin 1 was determined. On the basis of the purification data, it appears that interleukin 1 is biologically active within the 10(-11) to 10(-10) M concentration range. The success of the purification was due in part to the use of a superinduction protocol that resulted in the production of interleukin 1 at a rate of approximately 1 microgram/24 hr/4 X 10(6) P388D1 cells.