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垂体成纤维细胞生长因子:部分纯化与特性鉴定

Pituitary fibroblast growth factors: partial purification and characterization.

作者信息

Gambarini A G, Armelin H A

出版信息

Braz J Med Biol Res. 1981 Apr;14(1):19-27.

PMID:6458342
Abstract
  1. Isoelectric focusing of fractions derived from pH 7.0 extracts of bovine pituitary glands has been used to separate acidic and basic fibroblast growth factor (FGD) activity. FGF activity was assayed by measuring the initiation of DNA synthesis in Swiss mouse 3T3 fibroblasts. 2. Acidic and basic FGF activity present in crude extracts were not separated by chromatography on CM-Sephadex presumably because of association between acidic FGF and basic proteins. 3. Isoelectric focusing was used to separate acidic (pl 4-5) and basic (pI 8-9)FGF. The activity of acidic FGF was stable in urea and beta-mercaptoethanol, whereas basic FGF was inactivated. The activity of acidic FGF precipitated at pH 4.5 and was recovered when the precipitate was solubilized at pH 8, whereas basic FGF was soluble and stable at pH 4.5. 4. Acidic FGF at 10 ng/ml was as active as 1% fetal calf serum for the initiation of DNA synthesis. This potency is comparable with homogeneous preparations of basic FGF described in the literature.
摘要
  1. 利用等电聚焦法对从牛垂体pH 7.0提取物中获得的组分进行分离,以区分酸性和碱性成纤维细胞生长因子(FGF)活性。通过检测瑞士小鼠3T3成纤维细胞中DNA合成的起始情况来测定FGF活性。2. 粗提物中存在的酸性和碱性FGF活性在CM - 葡聚糖凝胶柱层析上未被分离,这可能是由于酸性FGF与碱性蛋白之间存在结合。3. 采用等电聚焦法分离酸性(pI 4 - 5)和碱性(pI 8 - 9)FGF。酸性FGF的活性在尿素和β - 巯基乙醇中稳定,而碱性FGF则失活。酸性FGF的活性在pH 4.5时沉淀,当沉淀在pH 8溶解时活性恢复,而碱性FGF在pH 4.5时可溶且稳定。4. 10 ng/ml的酸性FGF在引发DNA合成方面与1%胎牛血清的活性相当。这种效力与文献中描述的碱性FGF纯品制剂相当。

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