ABSTRACT

Background: The snake venom nerve growth factor (NGF)-induced signal transduction mechanism has never been explored.

Research design and methods: Homology modeling and molecular dynamic studies of the interaction between Russell’s viper venom NGF (RVV-NGFa) and mammalian tropomyosin-receptor kinase A (TrkA) was done by computational analysis. Transcriptomic and quantitative tandem mass spectrometry analyses determined the expression of intracellular genes and proteins, respectively, in RVV-NGFa-treated PC-12 neuronal cells. Small synthetic inhibitors of the signal transduction pathways were used to validate the major signaling cascades of neuritogenesis by RVV-NGFa.

Results: A comparative computational analysis predicted the binding of RVV-NGFa, mouse 2.5S-NGF (conventional neurotrophin), and Nn-α-elapitoxin-1 (non-conventional neurotrophin) to different domains of the TrkA receptor in PC-12 cells. The transcriptomic and quantitative proteomic analyses in unison showed differential expressions of common and unique genes and intracellular proteins, respectively, in RVV-NGFa-treated cells compared to control (untreated) mouse 2.5S-NGF and Nn-α-elapitoxin-1-treated PC-12 cells. The RVV-NGFa primarily triggered the mitogen-activated protein kinase-1 (MAPK1) signaling pathway for inducing neuritogenesis; however, 36 pathways of neuritogenesis were uniquely expressed in RVV-NGFa-treated PC-12 cells compared to mouse 2.5S NGF or Nn-α-elapitoxin-1 treated cells.

Conclusion: The common and unique intracellular signaling pathways of neuritogenesis by classical and non-classical neurotrophins were identified.

 抽象的

背景：蛇毒神经生长因子（NGF）诱导的信号转导机制从未被探索过。

研究设计和方法：通过计算分析对罗素毒蛇毒 NGF (RVV-NGFa) 和哺乳动物原肌球蛋白受体激酶 A (TrkA) 之间的相互作用进行同源建模和分子动力学研究。转录组和定量串联质谱分析分别测定了 RVV-NGFa 处理的 PC-12 神经元细胞中细胞内基因和蛋白质的表达。使用信号转导途径的小型合成抑制剂来验证 RVV-NGFa 神经突发生的主要信号级联。

结果：比较计算分析预测了 RVV-NGFa、小鼠 2.5S-NGF（传统神经营养蛋白）和 Nn-α-elapitoxin-1（非传统神经营养蛋白）与 PC-12 细胞中 TrkA 受体不同结构域的结合。转录组和定量蛋白质组分析一致显示，与对照（未处理）小鼠 2.5S-NGF 和 Nn-α-elapitoxin-1 处理的细胞相比，RVV-NGFa 处理的细胞中常见和独特基因以及细胞内蛋白质的表达存在差异。 PC-12 细胞。 RVV-NGFa 主要触发丝裂原激活蛋白激酶 1 (MAPK1) 信号通路，诱导神经突发生；然而，与小鼠 2.5S NGF 或 Nn-α-elapitoxin-1 处理的细胞相比，RVV-NGFa 处理的 PC-12 细胞中独特地表达了 36 条神经突发生途径。

结论：确定了经典和非经典神经营养因子神经突发生的常见和独特的细胞内信号传导途径。