Interactions between semaphorins and plexin-neuropilin receptor complexes in the membranes of live cells.
Interactions between semaphorins and plexin-neuropilin receptor complexes in the membranes of live cells.
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DOI：
10.1016/j.jbc.2021.100965
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发表时间：
2021-08
期刊：
影响因子：
--
通讯作者：
Smith AW
中科院分区：
文献类型：
--
作者：
Christie SM;Hao J;Tracy E;Buck M;Yu JS;Smith AW
信号蛋白配体通过其丛蛋白-神经毡蛋白受体的信号传导涉及发育中胚胎的组织模式。这些蛋白质在细胞迁移和粘附中发挥作用，但在疾病病因学中也很重要，包括癌症血管生成和转移。虽然这些受体的可溶性结构域的一些结构已经确定，但全长受体复合物的构象才刚刚开始被阐明，特别是在质膜的背景下。脉冲交错激发荧光互相关光谱可以直接洞察活细胞膜中蛋白质-蛋白质相互作用的形成。在这里，我们使用脉冲交错激发荧光互相关光谱研究了 Neuropilin-1 (Nrp1)、plexin A2、plexin A4 和 plexin D1 的同二聚化。与之前的研究一致，我们发现在没有外源配体的情况下，Nrp1、plexin A2 和 plexin A4 以二聚体形式存在。另一方面，Plexin D1 在相似的条件下是单体的，这在之前没有被报道过。我们还发现丛蛋白 A2 和 A4 组装成异聚复合物。当单独表达时，用信号蛋白 3A 或信号蛋白 3C 刺激既不会破坏也不会增强受体的二聚化，这表明激活涉及构象变化而不是单体-二聚体平衡的转变。然而，在信号蛋白 3C 刺激下，丛蛋白 D1 和 Nrp1 形成异聚复合物。这种相互作用的分析为现有的结构和生化数据提供了一种补充方法，将有助于开发针对癌症中这些受体的新治疗策略。
Signaling of semaphorin ligands via their plexin–neuropilin receptors is involved in tissue patterning in the developing embryo. These proteins play roles in cell migration and adhesion but are also important in disease etiology, including in cancer angiogenesis and metastasis. While some structures of the soluble domains of these receptors have been determined, the conformations of the full-length receptor complexes are just beginning to be elucidated, especially within the context of the plasma membrane. Pulsed-interleaved excitation fluorescence cross-correlation spectroscopy allows direct insight into the formation of protein–protein interactions in the membranes of live cells. Here, we investigated the homodimerization of neuropilin-1 (Nrp1), plexin A2, plexin A4, and plexin D1 using pulsed-interleaved excitation fluorescence cross-correlation spectroscopy. Consistent with previous studies, we found that Nrp1, plexin A2, and plexin A4 are present as dimers in the absence of exogenous ligand. Plexin D1, on the other hand, was monomeric under similar conditions, which had not been previously reported. We also found that plexin A2 and A4 assemble into a heteromeric complex. Stimulation with semaphorin 3A or semaphorin 3C neither disrupts nor enhances the dimerization of the receptors when expressed alone, suggesting that activation involves a conformational change rather than a shift in the monomer–dimer equilibrium. However, upon stimulation with semaphorin 3C, plexin D1 and Nrp1 form a heteromeric complex. This analysis of interactions provides a complementary approach to the existing structural and biochemical data that will aid in the development of new therapeutic strategies to target these receptors in cancer.
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影响因子：
4.7
作者：
Christie, Shaun M.;Ham, Trevor R.;Gilmore, Grant T.;Toth, Paul D.;Leipzig, Nic D.;Smith, Adam W.
通讯作者：
Smith, Adam W.
影响因子：
3.7
作者：
Gu, Chenghua;Giraudo, Enrico
通讯作者：
Giraudo, Enrico
影响因子：
8
作者：
Bagci, T.;Wu, J. K.;Jay, D. G.
通讯作者：
Jay, D. G.
影响因子：
11.8
作者：
Gitler, AD;Lu, MM;Epstein, JA
通讯作者：
Epstein, JA
DOI：
10.1007/978-1-4939-6448-2_1
发表时间：
2017
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Alto LT;Terman JR
通讯作者：
Terman JR