A novel 17 kD heparin-binding growth factor (HBGF-8) in bovine uterus: purification and N-terminal amino acid sequence.
A novel 17 kD heparin-binding growth factor (HBGF-8) in bovine uterus: purification and N-terminal amino acid sequence.
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DOI：
10.1016/0006-291x(89)92715-0
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发表时间：
1989-12
影响因子：
3.1
通讯作者：
Peter Gerard Milner;Yue-Sheng Li;R. M. Hoffman;Charles M. Kodner;Ned R. Siegel;Thomas F. Deuel
中科院分区：
文献类型：
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作者：
Peter Gerard Milner;Yue-Sheng Li;R. M. Hoffman;Charles M. Kodner;Ned R. Siegel;Thomas F. Deuel
我们从牛子宫中纯化了一种新的17 kD的生长因子，命名为肝素结合生长因子-8（HBGF-8）。生长因子与阳离子交换树脂和肝素-琼脂糖凝胶紧密结合，对丙酮沉淀稳定，在酸中不稳定。基于牛子宫丙酮提取物刺激血清饥饿的NIH 313细胞的~ 3 H-胸苷掺入DNA的总活性，使用丙酮粉末提取和中性pH下的色谱分离，HBGF-8的总活性达到6940倍纯化，总收率为0.4%。从1.2 kg湿重的组织中获得约18 μg蛋白。从17.5kD牛子宫碱性成纤维细胞生长因子（bFGF）中分离出HBGF-8，并对其N端氨基酸序列进行了分析。在小鼠NIH 3 T3成纤维细胞有丝分裂试验中，HBGF-8的活性与酸性成纤维细胞生长因子（aFGF）相当，略低于bFGF，在标准试验条件下，HBGF-8的内在比活性为5000 dpm/ng。
We have purified to near homogeneity a novel 17 kD growth factor from bovine uterus which we designated heparin-binding growth factor-8 (HBGF-8). The growth factor binds tightly to cation exchange resins and to Heparin-Sepharose and is stable to acetone precipitation and labile in acid. Based upon total activity in acetone extracts of bovine uterus stimulating3H-thymidine incorporation into DNA of serum-starved NIH 313 cells, a 6940 fold purification was achieved with an overall yield of HBGF-8 activity of 0.4%, using extraction of acetone powders and chromatographic separations at neutral pH. Approximately 18 μg protein was obtained from 1.2 kg wet weight of tissue. HBGF-8 was clearly separated from 17.5 kD bovine uterus basic fibroblast growth factor (bFGF) by purification and its N-terminal amino acid sequence analyzed. A polypeptide with a unique 25 N-terminal amino acid sequence was found. HBGF-8 was as active as acidic fibroblast growth factor (aFGF) and slightly less active than bFGF in the mouse NIH 3T3 fibroblast mitogenic assay system with an intrinsic specific activity of 5000 dpm/ng under standard assay conditions.