Spastin interacts with collapsin response mediator protein 3 to regulate neurite growth and branching.
Spastin interacts with collapsin response mediator protein 3 to regulate neurite growth and branching.
复制标题
Spastin 与塌陷素反应介导蛋白 3 相互作用，调节神经突生长和分支。
DOI：
10.4103/1673-5374.313052
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发表时间：
2021-12
影响因子：
6.1
通讯作者：
Lin HS
中科院分区：
文献类型：
--
作者：
Ji ZS;Li JP;Fu CH;Luo JX;Yang H;Zhang GW;Wu W;Lin HS
细胞骨架微管重排和运动对于脊髓损伤的修复至关重要。 Spastin 在微管切断的调节中发挥重要作用。 spastin和collapsin反应介导蛋白都可以调节神经突的生长和分支；然而，spastin在此过程中是否与崩溃素反应介导蛋白3（CRMP3）相互作用以及CRMP3参与脊髓损伤修复的机制尚不清楚。在这项研究中，我们使用蛋白质组学方法来鉴定与脊髓损伤修复相关的关键蛋白质。然后，我们采用液相色谱-质谱法来鉴定能够与谷胱甘肽 S-转移酶-spastin 相互作用的蛋白质。然后，使用免疫共沉淀和染色方法来评估 spastin 和 CRMP3 之间的潜在相互作用。最后，我们用 CRMP3 和 spastin 共转染原代海马神经元，以评估它们在神经突生长中的作用。质谱分析确定了 CRMP3 在脊髓损伤修复过程中的作用。液相色谱-质谱下拉分析鉴定出三种能够与 spastin 相互作用的 CRMP3 肽。 CRMP3 和 spastin 在脊髓中共表达，并且能够在体外和体内相互作用。最后，CRMP3 过表达能够增强 spastin 促进神经突生长和分支的能力。因此，我们的结果证实了 spastin 和 CRMP3 通过调节神经突生长和分支在脊髓损伤修复中发挥作用。因此，这些蛋白质可能是脊髓损伤修复的新靶点。中国暨南大学动物保育与使用委员会于2018年10月8日批准了本研究（批准号：IACUS-20181008-03）。
Cytoskeletal microtubule rearrangement and movement are crucial in the repair of spinal cord injury. Spastin plays an important role in the regulation of microtubule severing. Both spastin and collapsin response mediator proteins can regulate neurite growth and branching; however, whether spastin interacts with collapsin response mediator protein 3 (CRMP3) during this process remains unclear, as is the mechanism by which CRMP3 participates in the repair of spinal cord injury. In this study, we used a proteomics approach to identify key proteins associated with spinal cord injury repair. We then employed liquid chromatography-mass spectrometry to identify proteins that were able to interact with glutathione S-transferase-spastin. Then, co-immunoprecipitation and staining approaches were used to evaluate potential interactions between spastin and CRMP3. Finally, we co-transfected primary hippocampal neurons with CRMP3 and spastin to evaluate their role in neurite outgrowth. Mass spectrometry identified the role of CRMP3 in the spinal cord injury repair process. Liquid chromatography-mass spectrometry pulldown assays identified three CRMP3 peptides that were able to interact with spastin. CRMP3 and spastin were co-expressed in the spinal cord and were able to interact with one another in vitro and in vivo. Lastly, CRMP3 overexpression was able to enhance the ability of spastin to promote neurite growth and branching. Therefore, our results confirm that spastin and CRMP3 play roles in spinal cord injury repair by regulating neurite growth and branching. These proteins may therefore be novel targets for spinal cord injury repair. The Institutional Animal Care and Use Committee of Jinan University, China approved this study (approval No. IACUS-20181008-03) on October 8, 2018.