Purification and partial characterization of a megakaryocyte colony-stimulating factor from human plasma.
Purification and partial characterization of a megakaryocyte colony-stimulating factor from human plasma.
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从人血浆中纯化和部分表征巨核细胞集落刺激因子。
DOI：
10.1172/jci111813
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Straneva,JE
中科院分区：
文献类型：
--
作者：
Hoffman,R;Yang,HH;Bruno,E;Straneva,JE
从患有低巨核细胞性血小板减少症的患者获得的人血浆含有促进正常人骨髓细胞形成巨核细胞集落的因子。这种巨核细胞集落刺激因子是从这样的血浆标本中纯化的。经硫酸铵沉淀、二乙氨基乙基-Sepharose层析、麦胚凝集素-Sepharose 6 MB亲和层析和反相高效液相色谱四步纯化，其生物活性回收率为16.6%，比活性提高了3,489倍。纯化的蛋白在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上产生单一条带。纯化的巨核细胞集落刺激因子在浓度为7.6 × 10 ~（-8）M时能促进巨核细胞集落形成。巨核细胞集落刺激因子被证明是一种糖蛋白，具有明显的46，000 mol wt.巨核细胞集落刺激因子经三氟甲磺酸处理后去糖基化，导致其促进巨核细胞集落形成的能力丧失。巨核细胞集落刺激因子在巨核细胞集落形成单位水平上似乎是体外人巨核细胞生成的重要调节因子，可能具有重要的生理学意义。图片
Human plasma obtained from patients with hypomegakaryocytic thrombocytopenia contains a factor that promotes megakaryocyte colony formation by normal human marrow cells. This megakaryocyte colony-stimulating factor was purified from such a plasma specimen. A four-step purification scheme which included ammonium sulfate precipitation, diethylaminoethyl-Sepharose chromatography, affinity chromatography on wheat germ lectin-Sepharose 6MB, and reverse-phase high performance liquid chromatography resulted in a recovery of 16.6% of the initial biological activity and an increase in specific activity by 3,489-fold. The purified protein produced a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified megakaryocyte colony-stimulating factor was capable of promoting megakaryocyte colony formation at a concentration of 7.6 X 10(-8) M. Megakaryocyte colony-stimulating factor was shown to be a glycoprotein and had an apparent 46,000 mol wt. Deglycosylation of megakaryocyte colony-stimulating factor by treatment with trifluoromethane-sulfonate resulted in the loss of its ability to promote megakaryocyte colony formation. Megakaryocyte colony-stimulating factor appears to be an important regulator of in vitro human megakaryocytopoiesis at the level of the colony-forming unit megakaryocyte and may be of importance physiologically.Images