Analysis of Dispatched Protein Processing and Sonic Hedgehog Ligand Release.
Analysis of Dispatched Protein Processing and Sonic Hedgehog Ligand Release.
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DOI：
10.1007/978-1-0716-1701-4_9
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Ogden SK
中科院分区：
文献类型：
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作者：
Cleverdon E;Stewart DP;Ogden SK
12遍跨膜蛋白调度(DISP)是Sonic Hedgehog(SHH)从配体产生细胞释放所必需的，也是在组织构图过程中建立SHH形态原梯度所必需的。控制SHH释放的调控事件还没有完全确定。我们最近证明了DISP在其第一个胞外环的保守位置被Furin ProProtein Convertase切割。裂解位点的突变减弱了DISP介导的SHH的释放，这表明Furin裂解是促进DISP蛋白成熟的积极步骤。在本章中，我们提出了一种配体释放/保留方案，该方案允许分析DISP裂解、DISP介导的SHH配体从产生细胞中的释放以及靶细胞中分泌依赖的信号诱导。
The 12-pass transmembrane protein Dispatched (DISP) is essential for Sonic Hedgehog (SHH) release from ligand-producing cells and is indispensable for establishment of the SHH morphogen gradient during tissue patterning. Regulatory events controlling DISP release of SHH are not yet fully characterized. We recently demonstrated that DISP is cleaved by FURIN proprotein convertase at a conserved site in its first extracellular loop. Mutation of the cleavage site attenuates DISP-mediated SHH release, which indicates that Furin cleavage is a positive step toward DISP protein maturation. In this chapter, we present a ligand release/retention protocol that allows for the analysis of DISP cleavage, DISP-mediated release of SHH ligand from producing cells, and secretion-dependent signal induction in target cells.