Hansen Raino Kristian, Christensen Claus, Korshunova Irina, Kriebel Martin, Burkarth Nadine, Kiselyov Vladislav V, Olsen Marianne, Ostergaard Søren, Holm Arne, Volkmer Hansjürgen, Walmod Peter S, Berezin Vladimir, Bock Elisabeth
Protein Laboratory, Department of Neuroscience and Pharmacology, University of Copenhagen, Copenhagen, Denmark.
J Neurochem. 2007 Nov;103(4):1396-407. doi: 10.1111/j.1471-4159.2007.04894.x. Epub 2007 Sep 13.
A combinatorial library of undecapeptides was produced and utilized for the isolation of peptide binding to the fibronectin type 3 modules (F3I-F3II) of the neural cell adhesion molecule (NCAM). The isolated peptides were sequenced and produced as dendrimers. Two of the peptides (denoted ENFIN2 and ENFIN11) were confirmed to bind to F3I-F3II of NCAM by surface plasmon resonance. The peptides induced neurite outgrowth in primary cerebellar neurons and PC12E2 cells, but had no apparent neuroprotective properties. NCAM is known to activate different intracellular pathways, including signaling through the fibroblast growth factor receptor, the Src-related non-receptor tyrosine kinase Fyn, and heterotrimeric G-proteins. Interestingly, neurite outgrowth stimulated by ENFIN2 and ENFIN11 was independent of signaling through fibroblast growth factor receptor and Fyn, but could be inhibited with pertussis toxin, an inhibitor of certain heterotrimeric G-proteins. Neurite outgrowth induced by trans-homophilic NCAM was unaffected by the peptides, whereas knockdown of NCAM completely abrogated ENFIN2- and ENFIN11-induced neuritogenesis. These observations suggest that ENFIN2 and ENFIN11 induce neurite outgrowth in an NCAM-dependent manner through G-protein-coupled signal transduction pathways. Thus, ENFIN2 and ENFIN11 may be valuable for exploring this particular type of NCAM-mediated signaling.
制备了一个十一肽组合文库，并用于分离与神经细胞黏附分子（NCAM）的纤连蛋白III型结构域（F3I-F3II）结合的肽。对分离得到的肽进行测序并制备成树枝状聚合物。其中两个肽（分别命名为ENFIN2和ENFIN11）通过表面等离子体共振被证实可与NCAM的F3I-F3II结合。这些肽可诱导原代小脑神经元和PC12E2细胞的神经突生长，但没有明显的神经保护特性。已知NCAM可激活不同的细胞内信号通路，包括通过成纤维细胞生长因子受体、Src相关非受体酪氨酸激酶Fyn以及异源三聚体G蛋白进行信号传导。有趣的是，ENFIN2和ENFIN11刺激的神经突生长独立于通过成纤维细胞生长因子受体和Fyn的信号传导，但可被百日咳毒素（一种某些异源三聚体G蛋白的抑制剂）抑制。由反式同源NCAM诱导的神经突生长不受这些肽的影响，而敲低NCAM则完全消除了ENFIN2和ENFIN11诱导的神经突形成。这些观察结果表明，ENFIN2和ENFIN11通过G蛋白偶联信号转导途径以NCAM依赖的方式诱导神经突生长。因此，ENFIN2和ENFIN11对于探索这种特定类型的NCAM介导的信号传导可能具有重要价值。
