Application of a Mutant Cell Library to Determine the Structure-Function Relationship of Heparan Sulfate in Facilitating FGF2-FGFR1 Signaling.

Application of a Mutant Cell Library to Determine the Structure-Function Relationship of Heparan Sulfate in Facilitating FGF2-FGFR1 Signaling.
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应用突变细胞库确定硫酸乙酰肝素促进 FGF2-FGFR1 信号传导的结构功能关系。

DOI:
10.1007/978-1-0716-1398-6_48
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发表时间:
2022
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Wang,Lianchun
Wang,Lianchun
中科院分区:
--
文献类型:
--
作者:
Faulkner,John;Song,Xuehong;Wang,Lianchun

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硫酸乙酰肝素(HS)是一种结构复杂的线性多糖,可调节多种生物功能。阐明 HS 的结构-功能关系一直具有挑战性。最近,我们通过系统删除细胞中单独或组合表达的 HS 基因,生成了 HS 突变小鼠肺内皮细胞文库。在这里,我们描述了使用突变细胞库确定 HS 在细胞表面 FGF2 结合水平和下游细胞内信号传导水平上调节 FGF2-FGFR1 信号传导的结构功能关系的实验程序。我们的结果表明,HS 需要严格定义的精细结构才能充当 FGF2-FGFR1 信号传导的共受体。
Heparan sulfate (HS) is a linear polysaccharide with complex structures and modulates a wide range of biological functions. Elucidating the structure–function relationship of HS has been challenging. Recently, we generated a HS mutant mouse lung endothelial cell library by systematic deletion of HS genes expressed in the cell individually or in their combination. Here, we describe the experimental procedure using the mutant cell library to determine the structure–function relationship of HS in the regulation of FGF2-FGFR1 signaling at the levels of cell surface FGF2 binding and the downstream intracellular signaling activation. Our results demonstrated that strictly defined fine structure is required for HS to act as a co-receptor for FGF2-FGFR1 signaling.
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