Ruta M, Burgess W, Givol D, Epstein J, Neiger N, Kaplow J, Crumley G, Dionne C, Jaye M, Schlessinger J
Rorer Biotechnology, Inc., King of Prussia, PA 19406.
Proc Natl Acad Sci U S A. 1989 Nov;86(22):8722-6. doi: 10.1073/pnas.86.22.8722.
We have previously isolated a human gene from an endothelial cell cDNA library encoding a putative tyrosine kinase; we have designated this gene the fms-like gene (FLG). To analyze the gene product(s) of FLG, we have generated rabbit polyclonal antibodies directed against a synthetic peptide from FLG and used it to immunoprecipitate biosynthetically labeled FLG protein from a variety of human cell lines. These antibodies specifically recognized glycoprotein(s) of 100, 120, and 135 kDa with protein cores of 90 and 110 kDa. Acidic fibroblast growth factor (aFGF) stimulated tyrosine kinase activity of FLG in vitro and in living cells, suggesting that FLG encodes the membrane receptor for aFGF. Further supporting evidence came from cross-linking experiments on intact cells with the covalent cross-linking agent disuccinimidyl suberate and 125I-labeled aFGF as a specific probe. The cross-linked 125I-labeled aFGF-aFGF receptor complex was specifically immunoprecipitated with FLG antipeptide antibodies. It appears, therefore, that the receptor(s) for aFGF is related to the FLG gene product.
我们先前从一个内皮细胞cDNA文库中分离出一个人类基因,该基因编码一种假定的酪氨酸激酶;我们将此基因命名为fms样基因(FLG)。为了分析FLG的基因产物,我们制备了针对FLG合成肽的兔多克隆抗体,并用于从多种人类细胞系中免疫沉淀生物合成标记的FLG蛋白。这些抗体特异性识别100、120和135 kDa的糖蛋白,其蛋白核心为90和110 kDa。酸性成纤维细胞生长因子(aFGF)在体外和活细胞中刺激了FLG的酪氨酸激酶活性,这表明FLG编码aFGF的膜受体。进一步的支持证据来自于用共价交联剂辛二酸二琥珀酰亚胺酯和125I标记的aFGF作为特异性探针,对完整细胞进行的交联实验。交联的125I标记的aFGF-aFGF受体复合物用FLG抗肽抗体进行特异性免疫沉淀。因此,似乎aFGF的受体与FLG基因产物相关。