Center for Reproductive Medicine, Affiliated Hospital 1 of Wenzhou Medical University, Wenzhou, 325000, Zhejiang, People's Republic of China.
Department of Histology & Embryology, School of Basic Medical Science, Wenzhou Medical University, Cha Shan University Town, No.1 Central North Road, Wenzhou, 325035, Zhejiang, People's Republic of China.
J Mol Histol. 2021 Jun;52(3):521-537. doi: 10.1007/s10735-021-09960-6. Epub 2021 Feb 8.
Nogo-A protein consists of two main extracellular domains: Nogo-66 (rat amino acid [aa] 1019-1083) and Nogo-A-Δ20 (extracellular, active 180 amino acid Nogo-A region), which serve as strong inhibitors of axon regeneration in the adult CNS (Central Nervous System). Although receptors S1PR2 and HSPGs have been identified as Nogo-A-Δ20 binding proteins, it remains at present elusive whether other receptors directly interacting with Nogo-A-Δ20 exist, and decrease cell death. On the other hand, the key roles of EphA4 in the regulation of glioblastoma, axon regeneration and NSCs (Neural Stem Cells) proliferation or differentiation are well understood, but little is known the relationship between EphA4 and Nogo-A-Δ20 in NSCs apoptosis. Thus, we aim to determine whether Nogo-A-Δ20 can bind to EphA4 and affect survival of NSCs. Here, we discover that EphA4, belonging to a member of erythropoietin-producing hepatocellular (Eph) receptors family, could be acting as a high affinity ligand for Nogo-A-Δ20. Trans-membrane protein of EphA4 is needed for Nogo-A-Δ20-triggered inhibition of NSCs apoptosis, which are mediated by balancing p38 inactivation and JNK MAPK pathway activation. Finally, we predict at the atomic level that essential residues Lys-205, Ile-190, Pro-194 in Nogo-A-Δ20 and EphA4 residues Gln-390, Asn-425, Pro-426 might play critical roles in Nogo-A-Δ20/EphA4 binding via molecular docking.
Nogo-A 蛋白由两个主要的细胞外结构域组成:Nogo-66(大鼠氨基酸[aa] 1019-1083)和 Nogo-A-Δ20(细胞外,活性 180 个氨基酸的 Nogo-A 区域),它们是成年中枢神经系统(CNS)中轴突再生的强抑制剂。虽然已经确定 S1PR2 和 HSPGs 是 Nogo-A-Δ20 的结合蛋白,但目前尚不清楚是否存在其他直接与 Nogo-A-Δ20 相互作用的受体,并减少细胞死亡。另一方面,EphA4 在调节神经胶质瘤、轴突再生和神经干细胞(NSC)增殖或分化中的关键作用已得到充分理解,但对于 EphA4 和 Nogo-A-Δ20 在 NSC 凋亡中的关系知之甚少。因此,我们旨在确定 Nogo-A-Δ20 是否可以与 EphA4 结合并影响 NSC 的存活。在这里,我们发现 EphA4 属于促红细胞生成素产生肝细胞(Eph)受体家族的成员,可以作为 Nogo-A-Δ20 的高亲和力配体。EphA4 的跨膜蛋白对于 Nogo-A-Δ20 触发的 NSC 凋亡抑制是必需的,这是通过平衡 p38 失活和 JNK MAPK 通路激活来介导的。最后,我们从原子水平预测,Nogo-A-Δ20 中的关键残基 Lys-205、Ile-190、Pro-194 和 EphA4 残基 Gln-390、Asn-425、Pro-426 可能通过分子对接在 Nogo-A-Δ20/EphA4 结合中发挥关键作用。