Identification and characterization of synthetic chondroitin-4-sulfate binding peptides in neuronal functions
Identification and characterization of synthetic chondroitin-4-sulfate binding peptides in neuronal functions
复制标题
合成 4-硫酸软骨素结合肽在神经元功能中的鉴定和表征
DOI：
10.1038/s41598-018-37685-2
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发表时间：
2019-01-31
影响因子：
4.6
通讯作者：
Schachner, Melitta
中科院分区：
文献类型：
--
作者：
Loers, Gabriele;Liao, Yonghong;Schachner, Melitta
硫酸软骨素蛋白聚糖（CSPG）在哺乳动物脊髓损伤后的胶质瘢痕中及其周围上调，已被认为是通过阻碍轴突再生/发芽和突触重排而促进功能恢复的关键抑制分子。CSPG介导的抑制主要与CSPG的糖胺聚糖链相关，并且4-硫酸软骨素（C4 S）是调节成体神经系统中轴突引导和生长的主要硫酸化结构。为了寻找中和C4 S抑制功能的分子，我们筛选了与C4 S结合的肽的噬菌体展示文库。从与C4 S结合的噬菌体克隆中，我们选择了三种肽用于进一步分析。我们观察到这些肽以浓度依赖性和可饱和的方式与C4 S结合，但不与6-硫酸软骨素、硫酸肝素或硫酸皮肤素结合，而乱序肽显示与C4 S的结合高度减少或不结合。C4 S-结合肽，但不是他们的混乱的对应物，当添加到小鼠小脑神经元和人神经母细胞瘤细胞的培养物中时，中和C4 S-和CSPG-包被的基板对细胞粘附，神经元迁移和神经突生长的抑制功能。这些结果表明，C4 S结合肽中和CSPG的几种抑制功能，表明它们可能有利于修复哺乳动物神经系统损伤。
Chondroitin sulfate proteoglycans (CSPGs), up-regulated in and around the glial scar after mammalian spinal cord injury, have been suggested to be key inhibitory molecules for functional recovery by impeding axonal regrowth/sprouting and synaptic rearrangements. CSPG-mediated inhibition is mainly associated with the glycosaminoglycan chains of CSPGs, and chondroitin-4-sulfate (C4S) is the predominant sulfated structure that regulates axonal guidance and growth in the adult nervous system. With the aim to find molecules that neutralize the inhibitory functions of C4S, we screened a phage display library for peptides binding to C4S. From the phage clones binding to C4S we selected three peptides for further analysis. We observed that these peptides bind to C4S, but not chondroitin-6-sulfate, heparin sulfate or dermatan sulfate, in a concentration-dependent and saturable manner, whereas the scrambled peptides showed highly reduced or no binding to C4S. The C4S-binding peptides, but not their scrambled counterparts, when added to cultures of mouse cerebellar neurons and human neuroblastoma cells, neutralized the inhibitory functions of the C4S- and CSPG-coated substrate on cell adhesion, neuronal migration and neurite outgrowth. These results indicate that the C4S-binding peptides neutralize several inhibitory functions of CSPGs, suggesting that they may be beneficial in repairing mammalian nervous system injuries.