PURIFICATION AND BIOCHEMICAL-CHARACTERIZATION OF HUMAN PLURIPOTENT HEMATOPOIETIC COLONY-STIMULATING FACTOR
PURIFICATION AND BIOCHEMICAL-CHARACTERIZATION OF HUMAN PLURIPOTENT HEMATOPOIETIC COLONY-STIMULATING FACTOR
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DOI：
10.1073/pnas.82.5.1526
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
MOORE, MAS
中科院分区：
文献类型：
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作者：
WELTE, K;PLATZER, E;MOORE, MAS
多能造血集落刺激因子（pluripotent CSF）是由人膀胱癌细胞系5637组成型产生的蛋白质，从含有低血清（0.2%胎牛血清）的条件培养基中纯化。采用硫酸铵沉淀、离子交换层析、凝胶过滤和反相高效液相色谱等方法对粗多糖进行纯化。纯化的蛋白质在NaDodSO 4 [十二烷基硫酸钠]/聚丙烯酰胺凝胶电泳中具有18，000的MW，通过Ag染色技术和通过从相应的凝胶切片洗脱生物活性，并且具有5.5的等电点。多能CSF支持人混合集落、粒细胞-巨噬细胞集落和早期红细胞集落的生长，并诱导人早幼粒细胞白血病细胞系HL-60和鼠粒单核细胞白血病细胞系WEHI-3B（D+）分化。纯化的多能CSF在粒细胞-巨噬细胞集落测定中的比活性为1.5 × 10 - 6。108单位/毫克蛋白质。
Pluripotent hematopoietic colony-stimulating factor (pluripotent CSF), a protein that is constitutively produced by the human bladder carcinoma cell line 5637, was purified from low serum (0.2% fetal calf serum)-containing conditioned medium. The purification involved sequential ammonium sulfate precipitation, ion-exchange chromatography, gel filtration and reversed-phase high-performance liquid chromatography. The purified protein has a MW of 18,000 in NaDodSO4 [sodium dodecyl sulfate]/polyacrylamide gel electrophoresis, both by the Ag staining technique and by elution of biological activity from a corresponding gel slice, and has an isoelectric point of 5.5. Pluripotent CSF supports the growth of human mixed colonies, granulocyte-macrophage colonies and early erythroid colonies and induces differentiation of the human promyelocytic leukemic cell line HL-60 and the murine myelomonocytic leukemic cell line WEHI-3B (D+). The specific activity of the purified pluripotent CSF in the granulocyte-macrophage colony assay is 1.5 .times. 108 units/mg of protein.