Purification of a factor inducing differentiation in murine myelomonocytic leukemia cells. Identification as granulocyte colony-stimulating factor.
Purification of a factor inducing differentiation in murine myelomonocytic leukemia cells. Identification as granulocyte colony-stimulating factor.
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DOI：
10.1016/s0021-9258(18)32158-6
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发表时间：
1983-07
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影响因子：
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通讯作者：
N. Nicola;D. Metcalf;M. Matsumoto;G. Johnson
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文献类型：
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作者：
N. Nicola;D. Metcalf;M. Matsumoto;G. Johnson
小鼠骨髓单核细胞白血病细胞系（WEHI-3B）中天然存在的终末分化诱导剂从注射细菌内毒素的小鼠肺的条件培养基中纯化至表观均一性。通过使用盐析色谱法、苯基-琼脂糖色谱法、Bio-Gel P-60在1 M乙酸中的凝胶过滤、苯基-硅胶柱上的反相高效液相色谱法和凝胶过滤柱上的高效液相色谱法的连续分级分离，将该因子纯化超过400，000倍。在前两个步骤中，分化诱导因子与已知的正常骨髓细胞增殖调节因子粒细胞-巨噬细胞集落刺激因子完全分离，但在所有剩余步骤中，它与不同的粒细胞特异性集落刺激因子共纯化。纯化的因子在十二烷基硫酸钠-聚丙烯酰胺凝胶上显示Mr = 24，000 - 25，000的单一蛋白条带，与分化诱导和粒细胞集落刺激活性一致。粒细胞特异性集落刺激因子在体外对WEHI-3B细胞和正常粒细胞祖细胞具有活性，其半最大活性浓度为3 × 10 ~（-12）M。
A naturally occurring inducer of terminal differentiation in a murine myelomonocytic leukemia cell line (WEHI-3B) was purified to apparent homogeneity from medium conditioned by lungs from mice injected with bacterial endotoxin. The factor was purified over 400,000-fold by sequential fractionation using salting out chromatography, chromatography on phenyl-Sepharose, gel filtration on Bio-Gel P-60 in 1 M acetic acid, reverse-phase high performance liquid chromatography on a phenyl-silica column, and high performance liquid chromatography on a gel filtration column. During the first two steps, the differentiation-inducing factor was separated completely from a known proliferative regulator for normal myeloid cells, granulocyte-macrophage colony-stimulating factor, but it co-purified through all remaining steps with a distinct granulocyte-specific colony-stimulating factor. The purified factor showed a single protein band of Mr = 24,000-25,000 on sodium dodecyl sulfate-polyacrylamide gels coincident with both differentiation-inducing and granulocyte colony-stimulating activity. The granulocyte-specific colony-stimulating factor was active on WEHI-3B cells and normal granulocytic progenitor cells in vitro at the same half-maximally active concentration of 3 X 10(-12) M.