Hering Thomas M, Beller Justin A, Calulot Christopher M, Snow Diane M
Spinal Cord and Brain Injury Research Center, University of Kentucky, Lexington, KY 40536, USA.
Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH 44106, USA.
Biology (Basel). 2020 Feb 12;9(2):29. doi: 10.3390/biology9020029.
The role of proteoglycans in the central nervous system (CNS) is a rapidly evolving field and has major implications in the field of CNS injury. Chondroitin sulfate proteoglycans (CSPGs) increase in abundance following damage to the spinal cord and inhibit neurite outgrowth. Major advances in understanding the interaction between outgrowing neurites and CSPGs has created a need for more robust and quantitative analyses to further our understanding of this interaction. We report the use of a high-throughput assay to determine the effect of various post-translational modifications of aggrecan upon neurite outgrowth from NS-1 cells (a PC12 cell line derivative). Aggrecan contains chondroitin sulfate, keratan sulfate, and N-linked oligosaccharides (N-glycans), each susceptible to removal through different enzymatic digestions. Using a sequential digestion approach, we found that chondroitin sulfate and N-glycans, but not keratan sulfate, contribute to inhibition of neurite outgrowth by substrate-bound aggrecan. For the first time, we have shown that N-linked oligosaccharides on aggrecan contribute to its inhibition of neuritogenesis.
蛋白聚糖在中枢神经系统（CNS）中的作用是一个快速发展的领域，并且在CNS损伤领域具有重要意义。硫酸软骨素蛋白聚糖（CSPGs）在脊髓损伤后丰度增加，并抑制神经突生长。在理解生长中的神经突与CSPGs之间相互作用方面取得的重大进展，使得需要更强大和定量的分析方法来进一步加深我们对这种相互作用的理解。我们报告了使用一种高通量测定法来确定聚集蛋白聚糖的各种翻译后修饰对NS-1细胞（一种PC12细胞系衍生物）神经突生长的影响。聚集蛋白聚糖含有硫酸软骨素、硫酸角质素和N-连接寡糖（N-聚糖），每种都可通过不同的酶消化去除。使用顺序消化方法，我们发现硫酸软骨素和N-聚糖，而不是硫酸角质素，会导致底物结合的聚集蛋白聚糖抑制神经突生长。我们首次表明，聚集蛋白聚糖上的N-连接寡糖有助于其对神经突发生的抑制作用。
