Childs C B, Proper J A, Tucker R F, Moses H L
Proc Natl Acad Sci U S A. 1982 Sep;79(17):5312-6. doi: 10.1073/pnas.79.17.5312.
High concentrations of fetal bovine serum induced colony formation in soft agar by anchorage-dependent, nontransformed mouse AKR-2B and rat NRK cells. The colony-stimulating activity in fetal bovine serum was precipitated by 45% saturated ammonium sulfate and migrated in molecular sieve chromatography as a single peak of activity in the 10,000-15,000 molecular weight range. The colony-stimulating activity was heat and acid stable and was destroyed by trypsin and dithiothreitol, indicating the activity is due to a polypeptide that requires disulfide bonds for biological activity. No competition for binding to the epidermal growth factor receptor was associated with the colony-stimulating activity. Isoelectric focusing revealed activity in the pI 4-5 range. The colony-stimulating activity in serum appeared to be of platelet origin because platelet-poor plasma and platelet-poor plasma-derived serum contained little activity, whereas acid/ethanol extracts of bovine and human platelets had potent colony-stimulating activity. Chromatography of platelet extracts on Bio-Gel P-60 revealed peaks of AKR-2B colony-stimulating activity in the 12,000 and 20,000 molecular weight ranges. The other biological and chemical properties of the platelet colony-stimulating activity were the same as those for the serum activity. The data indicate the presence in serum of a platelet-derived growth factor(s) with properties similar to those of the transforming growth factors.
高浓度胎牛血清可诱导贴壁依赖性、未转化的小鼠AKR - 2B细胞和大鼠NRK细胞在软琼脂中形成集落。胎牛血清中的集落刺激活性可被45%饱和度的硫酸铵沉淀,在分子筛层析中,其活性以单一峰的形式迁移,分子量范围在10,000 - 15,000之间。集落刺激活性对热和酸稳定,但可被胰蛋白酶和二硫苏糖醇破坏,这表明该活性是由一种需要二硫键来维持生物活性的多肽引起的。集落刺激活性与表皮生长因子受体的结合不存在竞争关系。等电聚焦显示活性在pI 4 - 5范围内。血清中的集落刺激活性似乎来源于血小板,因为血小板贫乏的血浆和血小板贫乏血浆衍生的血清几乎没有活性,而牛和人血小板的酸/乙醇提取物具有强大的集落刺激活性。血小板提取物在Bio - Gel P - 60上的层析显示,在分子量12,000和20,000范围内有AKR - 2B集落刺激活性峰。血小板集落刺激活性的其他生物学和化学特性与血清活性相同。数据表明血清中存在一种血小板衍生生长因子,其性质与转化生长因子相似。