Graduate School of Pharmaceutical Science, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan.
Graduate School of Medical Sciences and Medical School, Nagoya City University, Kawasumi-1, Mizuho-cho Mizuho-ku, Nagoya 467-8601, Japan.
Biomolecules. 2011 Dec 14;1(1):48-62. doi: 10.3390/biom1010048.
Glycosylation improves the solubility and stability of proteins, contributes to the structural integrity of protein functional sites, and mediates biomolecular recognition events involved in cell-cell communications and viral infections. The first step toward understanding the molecular mechanisms underlying these carbohydrate functionalities is a detailed characterization of glycan structures. Recently developed glycomic approaches have enabled comprehensive analyses of N-glycosylation profiles in a quantitative manner. However, there are only a few reports describing detailed O-glycosylation profiles primarily because of the lack of a widespread standard method to identify O-glycan structures. Here, we developed an HPLC mapping method for detailed identification of O-glycans including neutral, sialylated, and sulfated oligosaccharides. Furthermore, using this method, we were able to quantitatively identify isomeric products from an in vitro reaction catalyzed by N-acetylglucosamine-6O-sulfotransferases and obtain O-glycosylation profiles of serum IgA as a model glycoprotein.
糖基化提高了蛋白质的溶解性和稳定性，有助于蛋白质功能部位的结构完整性，并介导涉及细胞间通讯和病毒感染的生物分子识别事件。了解这些碳水化合物功能的分子机制的第一步是对聚糖结构进行详细表征。最近开发的糖组学方法能够以定量方式全面分析 N-糖基化谱。然而，由于缺乏广泛使用的标准方法来鉴定 O-聚糖结构，仅有少数报道描述了详细的 O-糖基化谱。在这里，我们开发了一种 HPLC 图谱分析方法，用于详细鉴定包括中性、唾液酸化和硫酸化寡糖在内的 O-聚糖。此外，使用这种方法，我们能够从 N-乙酰氨基葡萄糖-6-O-磺基转移酶催化的体外反应中定量鉴定出异构体产物，并获得作为模型糖蛋白的血清 IgA 的 O-糖基化谱。
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