Molecular basis for Jagged-1/Serrate ligand recognition by the Notch receptor.
Molecular basis for Jagged-1/Serrate ligand recognition by the Notch receptor.
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DOI：
10.1074/jbc.m112.428854
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发表时间：
2013-03-08
期刊：
影响因子：
--
通讯作者：
Handford PA
中科院分区：
文献类型：
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作者：
Whiteman P;de Madrid BH;Taylor P;Li D;Heslop R;Viticheep N;Tan JZ;Shimizu H;Callaghan J;Masiero M;Li JL;Banham AH;Harris AL;Lea SM;Redfield C;Baron M;Handford PA
背景：Notch识别Jagged/Serrate配体的位点未知。结果如下：参与EGF 12的中心β-折叠的分子内疏水相互作用的两个关键残基形成配体结合平台。结论：配体结合区邻近参与调节Notch活性的Fringe敏感残基。重要性：这些结果对于理解受体/配体识别、Notch通过O-糖基化的调节以及旁系同源物特异性抗体的开发具有意义。我们已经映射了锯齿/锯齿结合位点的特定残基内的第12 EGF结构域的人和果蝇Notch。参与疏水相互作用的两个关键残基提供了配体结合平台，并与调节Notch活性的Fringe敏感残基相邻。我们的数据表明，结合位点内的小变化微调配体特异性，这可以解释在哺乳动物Notch旁系同源物中观察到的序列异质性，并应允许开发旁系同源物特异性配体阻断抗体。作为原理证明，我们已经产生了一种阻断结合的Notch-1特异性单克隆抗体，从而为研究和治疗应用的抗体工具铺平了道路。
Background: The site of Jagged/Serrate ligand recognition by Notch is unknown. Results: Two critical residues involved in an intramolecular hydrophobic interaction across the central β-sheet of EGF12 form a ligand-binding platform. Conclusion: The ligand-binding region is adjacent to a Fringe-sensitive residue involved in modulating Notch activity. Significance: The results have implications for understanding receptor/ligand recognition, Notch regulation by O-glycosylation, and the development of paralogue-specific antibodies. We have mapped a Jagged/Serrate-binding site to specific residues within the 12th EGF domain of human and Drosophila Notch. Two critical residues, involved in a hydrophobic interaction, provide a ligand-binding platform and are adjacent to a Fringe-sensitive residue that modulates Notch activity. Our data suggest that small variations within the binding site fine-tune ligand specificity, which may explain the observed sequence heterogeneity in mammalian Notch paralogues, and should allow the development of paralogue-specific ligand-blocking antibodies. As a proof of principle, we have generated a Notch-1-specific monoclonal antibody that blocks binding, thus paving the way for antibody tools for research and therapeutic applications.