Ligand-affinity cloning and structure of a cell surface heparan sulfate proteoglycan that binds basic fibroblast growth factor.
Ligand-affinity cloning and structure of a cell surface heparan sulfate proteoglycan that binds basic fibroblast growth factor.
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DOI：
10.1073/pnas.87.18.6985
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发表时间：
1990-09
影响因子：
11.1
通讯作者：
M. Kiefer;J. Stephans;K. Crawford;K. Okino;P. Barr
中科院分区：
文献类型：
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作者：
M. Kiefer;J. Stephans;K. Crawford;K. Okino;P. Barr
编码碱性成纤维细胞生长因子（FGF）结合蛋白的cDNA的表达克隆证实了以前的假设，即该分子是细胞表面硫酸乙酰肝素蛋白聚糖。将从富含FGF受体活性的仓鼠肾细胞系构建的cDNA文库转染到人淋巴母细胞系中。在其表面表达功能性碱性FGF结合蛋白的克隆通过在用人碱性FGF包被的塑料皿上淘选来富集。从分离的cDNA推导的氨基酸序列揭示了几个有趣的功能，包括疏水信号和跨膜结构域，侧翼的胞外区含有6个潜在的附着位点的糖胺聚糖侧链。该结构还包含一个短的亲水性细胞质尾序列同源先前报道的肌动蛋白结合域。碱性FGF与表达结合蛋白的细胞的结合可被肝素和硫酸乙酰肝素抑制，但不被硫酸软骨素、硫酸皮肤素或硫酸角质素抑制。除了结合碱性FGF之外，该蛋白质或相关表面蛋白质还可以作为其它生长因子和病毒（如单纯疱疹病毒）的初始细胞附着位点。
Expression cloning of cDNAs encoding a basic fibroblast growth factor (FGF) binding protein confirms previous hypotheses that this molecule is a cell-surface heparan sulfate proteoglycan. A cDNA library constructed from a hamster kidney cell line rich in FGF receptor activity was transfected into a human lymphoblastoid cell line. Clones expressing functional basic FGF binding proteins at their surfaces were enriched by panning on plastic dishes coated with human basic FGF. The amino acid sequence deduced from the isolated cDNAs revealed several interesting features, including hydrophobic signal and transmembrane domains that flank an extracellular region containing six potential attachment sites for glycosaminoglycan side chains. The structure also contains a short hydrophilic cytoplasmic tail sequence homologous to previously reported actin binding domains. Binding of basic FGF to cells expressing the binding protein could be inhibited by heparin and heparan sulfate but not by chondroitin sulfate, dermatan sulfate, or keratan sulfate. In addition to binding basic FGF, this protein or related surface proteins may function as an initial cellular attachment site for other growth factors and for viruses, such as herpes simplex virus.