GlycoFly: A Database of Drosophila N-linked Glycoproteins Identified Using SPEG-MS Techniques
GlycoFly: A Database of Drosophila N-linked Glycoproteins Identified Using SPEG-MS Techniques
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DOI：
10.1021/pr200004t
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发表时间：
2011-06-01
影响因子：
4.4
通讯作者：
Betenbaugh, Michael J.
中科院分区：
文献类型：
--
作者：
Baycin-Hizal, Deniz;Tian, Yuan;Betenbaugh, Michael J.
蛋白质糖基化影响中枢神经系统（CNS）的细胞功能。它的缺乏会导致神经系统疾病，如共济失调、瘫痪、学习障碍、智力低下和记忆丧失。然而，导致这些疾病的糖蛋白尚未得到很好的表征。本研究以果蝇为模式生物，通过蛋白质组学手段鉴定其中枢神经系统的N-糖基化蛋白及其N-糖基化位点。用胰凝乳蛋白酶或胰蛋白酶消化成蝇头，并使用 N-连接糖肽固相萃取 (SPEG) 技术捕获 N-连接糖肽，然后使用 LTQ OrbiTrap Velos 进行质谱 (MS) 分析。从 721 个独特检测到的具有 740 个 NXS/T 糖基化位点的肽中鉴定出 330 种新糖蛋白和 147 种先前已知的糖蛋白。 N-糖基化位点在细胞粘附、离子通道和离子结合分子中非常丰富，这对于神经成熟、器官发育、轴突引导、学习和记忆非常重要。这些蛋白质的 N-糖基化位点的鉴定将增强我们对这些蛋白质的了解，并为未来研究这些蛋白质在神经功能和疾病中的作用奠定基础。本研究建立了果蝇 N 连接糖肽数据库 (http://betenbaugh.jhu.edu/GlycoFly)，作为神经系统疾病研究的资源。
Protein glycosylation affects cellular functions of the central nervous system (CNS). Its deficiency leads to neurological disorders such as ataxia, paralysis, learning disability, mental retardation, and memory loss. However, the glycoproteins that are responsible for these diseases are not well characterized. In this study, Drosophila melanogaster was used as a model organism to identify the N-glycosylated proteins and N-glycosylation sites of its CNS by means of proteomics. Adult fly heads were digested with chymotrypsin or trypsin and the N-linked glycopeptides were captured using solid phase extraction of N-linked glycopeptides (SPEG) technique followed by mass spectrometry (MS) analysis using LTQ OrbiTrap Velos. Three hundred and thirty new and 147 previously known glycoproteins were identified from 721 uniquely detected peptides that have 740 NXS/T glycosylation sites. The N-glycosylation sites were highly abundant in cell adhesion, ion channel, and ion binding molecules, which are important for nerve maturation, organ development, axon guidance, learning, and memory. Identification of the N-glycosylated sites of these proteins will enhance our knowledge of these proteins and serve as a basis for future studies to address the roles of these proteins in neurological function and disorders. A database for Drosophila N-linked glycopeptides (http://betenbaugh.jhu.edu/GlycoFly) has been established in this study as a resource for study of neurological disorders.