IDENTIFICATION OF THE FIBROBLAST GROWTH-FACTOR RECEPTOR OF SWISS-3T3 CELLS AND MOUSE SKELETAL-MUSCLE MYOBLASTS
IDENTIFICATION OF THE FIBROBLAST GROWTH-FACTOR RECEPTOR OF SWISS-3T3 CELLS AND MOUSE SKELETAL-MUSCLE MYOBLASTS
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DOI：
10.1021/bi00360a001
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发表时间：
1986-06-17
期刊：
影响因子：
2.9
通讯作者：
HAUSCHKA, SD
中科院分区：
文献类型：
--
作者：
OLWIN, BB;HAUSCHKA, SD
根据两种不同的成纤维细胞生长因子(FGF)刺激骨骼肌成肌细胞增殖的能力，从牛脑中提纯得到两种不同的成纤维细胞生长因子。这些生长因子对瑞士3T3细胞也是有丝分裂的，似乎与之前分离的阴离子和阳离子成纤维细胞生长因子密切相关或相同。阴阳离子生长因子刺激成肌细胞DNA合成的半数浓度(EC50)分别为30 pM和1 pM。相反，两种生长因子刺激3T3细胞DNA合成的EC50均为45 pm。~(125)I标记的阴离子成纤维细胞生长因子的结合是饱和的，表观Kd值分别为45 pm和11 pm，3T3细胞和MM14小鼠成肌细胞的Kd值分别约为60,000和2000个/细胞。未标记的阴离子和阳离子的成纤维细胞生长因子平均取代3T3细胞中的125I标记的阴离子成纤维细胞生长因子，而阳离子成纤维细胞生长因子比阴离子成肌细胞的置换能力更强，表明这两种不同的生长因子是由单一的受体结合的。由于血小板衍生生长因子、胰岛素、胰岛素样生长因子1、表皮生长因子和神经生长因子不能取代瑞士3T3细胞中结合的125I标记的阴离子成纤维细胞生长因子，因此这些因子的结合是特异的。将125I标记的阴离子成纤维细胞生长因子与3T3细胞和MM14成肌细胞进行化学交联，鉴定出单一的洗涤剂可溶成纤维细胞生长因子受体，其表观分子量为165,000。
Two distinct fibroblast growth factors (FGF) were purified to homogeneity from bovine brain on the basis of their ability to stimulate skeletal muscle myoblast proliferation. These growth factors are also mitogenic for Swiss 3T3 cells and appear to be closely related to or identical with previously isolated anionic and cationic fibroblast growth factors. The half-maximum concentrations (EC50) for stimulation of myoblast DNA synthesis by the anionic and cationic growth factors were 30 pM and 1 pM, respectively. In contrast, an EC50 of 45 pM was observed for stimulation of 3T3 cell DNA synthesis by both growth factors. Binding of 125I-labeled anionic FGF was saturable with apparent Kd values of 45 pM and 11 pM and approximately 60,000 and 2000 receptor sites per cell for 3T3 cells and MM14 murine myoblasts, respectively. Unlabeled anionic and cationic FGF equally displaced 125I-labeled anionic FGF from 3T3 cells while cationic FGF was more potent than anionic FGF for displacement from skeletal muscle myoblasts, demonstrating that a single receptor binds the two distinct growth factors. Binding was specific for these factors since platelet-derived growth factor, insulin, insulin-like growth factor 1, epidermal growth factor, and nerve growth factor were unable to displace bound 125I-labeled anionic FGF from Swiss 3T3 cells. Chemical cross-linking of specifically bound 125I-labeled anionic FGF to 3T3 cells and MM14 myoblasts identified a single detergent-soluble FGF receptor with an apparent molecular weight of 165,000.