Reelin (400 kDa) is an extracellular matrix glycoprotein that is a key regulator of the many significant biological processes including the brain formation, cell aggregation, and dendrite formation. The glycosylation contributes to the nature of the protein through folding, localization and trafficking, solubility, antigenicity, biological activity, and half-life. Although reelin is to be known as a glycoprotein, the knowledge of its glycosylation is very limited. In this study, we aimed to characterize the terminal glycan profile of reelin by lectin blotting and monosaccharide analysis of glycan chains by capillary liquid chromatography electrospray ionization ion trap tandem mass spectrometry (CapLC-ESI-MS/MS) in SH-SY5Y neuroblastoma cell line. According to our results, reelin was detected in different protein fragments (310, 250, and 85 kDa) in addition to full-length form (400 kDa) in the cell line. The reelin glycoprotein was found to carry the β-N-Acetylglucosamine, α-Mannose, β-Galactose, and α-2,3 and α2,6 linked sialic acids by lectin blotting. Nevertheless, these terminal monosaccharides were found in different intensity according to reelin fragments. Besides, we purified a reelin fragment (250 kDa), and we analyzed it for their monosaccharide by CapLC-ESI-MS/MS. We found that reelin contained five types of monosaccharides, which were consisted of N-Acetylgalactosamine, N-Acetylglucosamine, Galactose, Glucose, Mannose and Sialic acid, from high to low abundance respectively. The present results provide a valuable guide for biochemical, genetic, and glycobiology based further experiments about reelin glycosylation in cancer perspective.

Reelin（400 kDa）是一种细胞外基质糖蛋白，是许多重要生物过程（包括脑形成，细胞聚集和树突形成）的关键调节剂。糖基化通过折叠，定位和运输，溶解性，抗原性，生物活性和半衰期而有助于蛋白质的性质。尽管瑞林被称为糖蛋白，但是其糖基化的知识非常有限。在这项研究中，我们旨在通过毛细管液色谱电喷雾电离离子阱串联质谱法（CapLC-ESI-MS / MS）在SH-SY5Y神经母细胞瘤细胞系中通过凝集素印迹和聚糖链的单糖分析来表征reelin的末端聚糖谱。 。根据我们的结果，在不同的蛋白质片段中检测到了reelin（310、250，细胞系中的全长形式（400 kDa）和85 kDa）。通过凝集素印迹，发现reelin糖蛋白携带β-N-乙酰基葡糖胺，α-甘露糖，β-半乳糖以及α-2,3和α2,6连接的唾液酸。然而，根据reelin片段，发现这些末端单糖的强度不同。此外，我们纯化了一个reelin片段（250 kDa），并通过CapLC-ESI-MS / MS分析了它们的单糖。我们发现reelin包含五种类型的单糖，分别由高丰度到低丰度分别由N-乙酰半乳糖胺，N-乙酰氨基葡萄糖，半乳糖，葡萄糖，甘露糖和唾液酸组成。本研究结果为基于瑞兰糖基化的进一步实验从生物化学，遗传和糖生物学提供了有价值的指导。