GlycoFish: a database of zebrafish N-linked glycoproteins identified using SPEG method coupled with LC/MS.
GlycoFish: a database of zebrafish N-linked glycoproteins identified using SPEG method coupled with LC/MS.
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DOI：
10.1021/ac200726q
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发表时间：
2011-07-01
影响因子：
7.4
通讯作者：
Zhang, Hui
中科院分区：
文献类型：
--
作者：
Baycin-Hizal, Deniz;Tian, Yuan;Akan, Ilhan;Jacobson, Elena;Clark, Dean;Wu, Alexander;Jampol, Russell;Palter, Karen;Betenbaugh, Michael;Zhang, Hui
斑马鱼(Danio Rerio)是研究人类疾病机制和途径的模式生物。许多神经系统、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建立。本研究以斑马鱼胚胎糖蛋白质组学为基础，研究了斑马鱼胚胎中N-连接的糖蛋白及其糖基化位点。为了增加斑马鱼糖肽的数量，用两种不同的酶--胰凝乳酶和胰蛋白酶将斑马鱼蛋白质消化成不同长度的多肽。采用固相萃取N-连接糖肽(SpeG)的方法提取斑马鱼的N-糖基化多肽，并用LTQ Orbitrap Velos质谱仪进行鉴定。从265个独特的糖肽中，包括269个NXT/S糖位点，我们鉴定了169个不同的N-糖基化蛋白。已鉴定的糖蛋白在转运蛋白、细胞黏附和离子通道/离子结合类蛋白中含量非常丰富，这些蛋白对胚胎、器官和中枢神经系统的发育至关重要。这些蛋白质组学数据将扩大我们对斑马鱼糖蛋白的了解，并可能用于阐明糖基化在细胞过程和疾病中所起的作用。糖蛋白数据可通过本文介绍的GlycoFish数据库(http://betenbaugh.jhu.edu/GlycoFish))获得。
Zebrafish (Danio rerio) is a model organism 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 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.
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