Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA.
Anal Chem. 2011 Jul 1;83(13):5296-303. doi: 10.1021/ac200726q. Epub 2011 Jun 8.
Zebrafish (Danio rerio) is a model organism that is used to study the mechanisms and pathways of human disorders. Many dysfunctions in neurological, development, and neuromuscular systems are due to glycosylation deficiencies, but the glycoproteins involved in zebrafish embryonic development have not been established. In this study, a mass spectrometry-based glycoproteomic characterization of zebrafish embryos was performed to identify the N-linked glycoproteins and N-linked glycosylation sites. To increase the number of glycopeptides, proteins from zebrafish were digested with two different proteases--chymotrypsin and trypsin--into peptides of different length. The N-glycosylated peptides of zebrafish were then captured by the solid-phase extraction of N-linked glycopeptides (SPEG) method and the peptides were identified with an LTQ OrbiTrap Velos mass spectrometer. From 265 unique glycopeptides, including 269 consensus NXT/S glycosites, we identified 169 different N-glycosylated proteins. The identified glycoproteins were highly abundant in proteins belonging to the transporter, cell adhesion, and ion channel/ion binding categories, which are important to embryonic, organ, and central nervous system development. This proteomics data will expand our knowledge about glycoproteins in zebrafish and may be used to elucidate the role that glycosylation plays in cellular processes and disease. The glycoprotein data are available through the GlycoFish database (http://betenbaugh.jhu.edu/GlycoFish) introduced in this paper.
斑马鱼（Danio rerio）是一种模式生物，用于研究人类疾病的机制和途径。许多神经、发育和神经肌肉系统的功能障碍是由于糖基化缺陷引起的，但参与斑马鱼胚胎发育的糖蛋白尚未确定。在这项研究中，我们进行了基于质谱的斑马鱼胚胎糖蛋白质组学特征分析，以鉴定 N-连接糖蛋白和 N-连接糖基化位点。为了增加糖肽的数量，我们使用两种不同的蛋白酶——胰凝乳蛋白酶和胰蛋白酶——将斑马鱼的蛋白质消化成不同长度的肽。然后，通过固相萃取 N-连接糖肽（SPEG）方法捕获斑马鱼的 N-糖肽，并用 LTQ OrbiTrap Velos 质谱仪鉴定肽。从 265 个独特的糖肽中，包括 269 个共识 NXT/S 糖基化位点，我们鉴定了 169 种不同的 N-糖基化蛋白。鉴定出的糖蛋白在属于转运体、细胞黏附和离子通道/离子结合类别的蛋白质中含量丰富，这些蛋白质对胚胎、器官和中枢神经系统的发育很重要。这些蛋白质组学数据将扩展我们对斑马鱼糖蛋白的认识，并可用于阐明糖基化在细胞过程和疾病中的作用。糖蛋白数据可通过本文介绍的 GlycoFish 数据库（http://betenbaugh.jhu.edu/GlycoFish）获得。
