Processed Sonic hedgehog - an active signaling protease?

加工过的 Sonic Hedgehog - 一种活性信号蛋白酶？

• 批准号: 246417003
• 负责人: Professor Dr. Kay Grobe
• 金额: --
• 依托单位: Institut für Physiologische Chemie und Pathobiochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/246417003?language=en
• 关键词: Processed; Sonic; hedgehog; active; signaling

A major challenge in developmental biology is to understand how cells coordinate developmental behaviors with that of their neighbours. Cells often employ secreted signaling molecules such as the Hedgehog (Hh) morphogens to control developmental growth and patterning. Hh is an unusual signaling molecule, however. All vertebrate Hh family members are metalloproteins (containing a tetrahedrally coordinated zinc ion) that are synthesized in dually lipid-modified form (N-terminally palmitoylated, C-terminally cholesterol-linked). This results in metalloprotein multimerization and firm tethering to the surface of producing cells. Yet, Hhs do get released from producing cells and form lipidation-dependent concentration gradients. This raises the question of how Hh release is achieved, and specifically, how Hh release is linked to morphogen gradient formation. Previously, we showed that N-terminal lipidation is the prerequisite for the processing of an associated N-terminal peptide during sheddase-mediated Sonic Hh (Shh) release from producing cells. In addition to Shh solubilization, removal of N-terminal peptides results in the exposure of the Shh tetrahedral zinc-coordination site, which serves as the receptor (Patched, Ptc) binding site. This site was previously blocked by the N-terminal peptide. Importantly, the Shh zinc-coordination site and lysostaphin-type protease active sites are strikingly similar, indicating that Shh bears a functional protease active site. In this grant proposal, we thus suggest that N-terminal peptide processing during release is coupled to the conversion of an inactive, membrane-tethered Shh zymogen into an active zinc-metalloprotease. By processing extracellular matrix constituents, and possibly via autodegradation, Shh proteolytic activation may then help shape its extracellular gradient. To test this idea, we aim to characterize in vitro and in vivo Shh targets and specificity via modern, system-wide protease substrate profiling technology. We also plan to test putative roles of functional Hh zinc-coordination sites in the well-characterized Drosophila system in vivo. From these experiments, we expect fascinating insights into proteolysis-dependent Hh gradient formation in vivo and in vitro, with important implications for development and disease.

发育生物学的一个主要挑战是了解细胞如何与其邻近细胞协调发育行为。细胞经常利用分泌的信号分子，例如刺猬 (Hh) 形态发生素来控制发育生长和模式形成。然而，Hh 是一种不寻常的信号分子。所有脊椎动物 Hh 家族成员都是金属蛋白（含有四面体配位的锌离子），以双重脂质修饰形式（N 末端棕榈酰化、C 末端胆固醇连接）合成。这导致金属蛋白多聚化并牢固地束缚在生产细胞的表面。然而，Hhs 确实从生产细胞中释放出来，并形成脂化依赖性浓度梯度。这就提出了如何实现 Hh 释放的问题，具体来说，Hh 释放如何与形态发生素梯度形成相关。之前，我们表明 N 端脂化是在脱落酶介导的 Sonic Hh (Shh) 从生产细胞释放过程中加工相关 N 端肽的先决条件。除了 Shh 溶解之外，去除 N 端肽还会导致 Shh 四面体锌配位位点暴露，该位点充当受体（Patched，Ptc）结合位点。该位点之前被 N 端肽封闭。重要的是，Shh 锌配位位点和溶葡萄球菌素型蛋白酶活性位点惊人相似，表明 Shh 具有功能性蛋白酶活性位点。因此，在这项资助提案中，我们建议释放过程中的 N 端肽加工与无活性、膜束缚的 Shh 酶原转化为活性锌金属蛋白酶相结合。通过处理细胞外基质成分，并可能通过自降解，Shh 蛋白水解激活可能有助于塑造其细胞外梯度。为了测试这个想法，我们的目标是通过现代全系统蛋白酶底物分析技术来表征体外和体内 Shh 靶点和特异性。我们还计划测试功能性 Hh 锌配位位点在充分表征的果蝇体内系统中的假定作用。从这些实验中，我们期望能够深入了解体内和体外蛋白水解依赖性 Hh 梯度的形成，这对发育和疾病具有重要意义。

项目成果

Ca2+ coordination controls sonic hedgehog structure and its Scube2-regulated release

• DOI: 10.1242/jcs.205872
• 发表时间: 2017-10-01
• 期刊: JOURNAL OF CELL SCIENCE
• 影响因子: 4
• 作者: Jakobs, Petra;Schulz, Philipp;Grobe, Kay
• 通讯作者: Grobe, Kay

Soluble Heparin and Heparan Sulfate Glycosaminoglycans Interfere with Sonic Hedgehog Solubilization and Receptor Binding

• DOI: 10.3390/molecules24081607
• 发表时间: 2019-04-02
• 期刊: MOLECULES
• 影响因子: 4.6
• 作者: Manikowski, Dominique;Jakobs, Petra;Grobe, Kay
• 通讯作者: Grobe, Kay

Disrupting Hedgehog Cardin–Weintraub sequence and positioning changes cellular differentiation and compartmentalization in vivo

• DOI: 10.1242/dev.167221
• 发表时间: 2018-09
• 期刊: Development
• 影响因子: 4.6
• 作者: Philipp Kastl;D. Manikowski;Georg Steffes;S. Schürmann;Shyam Bandari;C. Klämbt;K. Grobe