Purification and partial amino acid sequence of asialo murine granulocyte-macrophage colony stimulating factor.
Purification and partial amino acid sequence of asialo murine granulocyte-macrophage colony stimulating factor.
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去唾液酸鼠粒细胞-巨噬细胞集落刺激因子的纯化和部分氨基酸序列。
DOI：
10.1073/pnas.82.2.292
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发表时间：
1985
影响因子：
11.1
通讯作者：
Burgess,AW
中科院分区：
文献类型：
--
作者：
Sparrow,LG;Metcalf,D;Hunkapiller,MW;Hood,LE;Burgess,AW
建立了一种从小鼠肺条件培养液中分离纯化粒细胞-巨噬细胞集落刺激因子(asialo-GM-CSF)的方法。在纯化中，部分纯化的因子用神经氨酸酶处理，以减少由于唾液酸化程度不同而导致的电荷不均一性。最后的反相液相色谱步骤分离了三种活性形式的阿西洛因子。在聚丙烯酰胺凝胶电泳法上，它们各有一条主带和几条次带，具有相似的氨基酸组成。纯化的小鼠asialo-GM-CSF的比活性约为每毫克蛋白8×10(9)个菌落。主要形式的氨基酸序列测定给出了一个单一的氨基末端序列，该序列已被用于制备寡核苷酸探针，用于分离两个编码GM-CSF的cDNA克隆。这两个克隆的核苷酸序列推导出的氨基酸序列与asialo-GM-CSF的氨基酸末端几乎相同，氨基酸组成与asialo-GM-CSF非常相似。
A procedure utilizing reversed-phase high-performance liquid chromatography is described for the purification of asialo granulocyte-macrophage colony stimulating factor (asialo-GM-CSF) from mouse lung-conditioned medium. In the purification, the partially purified factor was treated with neuraminidase to reduce charge heterogeneity due to variable degrees of sialation. Three active forms of the asialo factor were separated by the final reversed-phase liquid chromatography step. These each gave a single major band and several minor bands on polyacrylamide gel electrophoresis and had similar amino acid compositions. The specific activity of purified murine asialo-GM-CSF was approximately 8 X 10(9) colonies per mg of protein. Amino acid sequence determination of the major form gave a single amino-terminal sequence, which has been used to develop oligonucleotide probes for the isolation of two cDNA clones encoding GM-CSF. The nucleotide sequence of these two clones gave a deduced amino acid sequence almost identical with that determined for the amino terminus of asialo-GM-CSF and an amino acid composition very similar to that for asialo-GM-CSF.
生长因子和干细胞
DOI：
--
发表时间：
1984
期刊：
影响因子：
--
作者：
A. Burgess;N. Nicola
通讯作者：
N. Nicola