Magnelli Paula, Cipollo John F, Ratner Daniel M, Cui Jike, Kelleher Daniel, Gilmore Reid, Costello Catherine E, Robbins Phillips W, Samuelson John
Department of Molecular and Cell Biology, Department of Biochemistry, Boston University Medical Center, Boston, MA 02118-2526, USA.
J Biol Chem. 2008 Jun 27;283(26):18355-64. doi: 10.1074/jbc.M800725200. Epub 2008 Apr 16.
N-Glycans of Entamoeba histolytica, the protist that causes amebic dysentery and liver abscess, are of great interest for multiple reasons. E. histolytica makes an unusual truncated N-glycan precursor (Man(5)GlcNAc(2)), has few nucleotide sugar transporters, and has a surface that is capped by the lectin concanavalin A. Here, biochemical and mass spectrometric methods were used to examine N-glycan biosynthesis and the final N-glycans of E. histolytica with the following conclusions. Unprocessed Man(5)GlcNAc(2), which is the most abundant E. histolytica N-glycan, is aggregated into caps on the surface of E. histolytica by the N-glycan-specific, anti-retroviral lectin cyanovirin-N. Glc(1)Man(5)GlcNAc(2), which is made by a UDP-Glc: glycoprotein glucosyltransferase that is part of a conserved N-glycan-dependent endoplasmic reticulum quality control system for protein folding, is also present in mature N-glycans. A swainsonine-sensitive alpha-mannosidase trims some N-glycans to biantennary Man(3)GlcNAc(2). Complex N-glycans of E. histolytica are made by the addition of alpha1,2-linked Gal to both arms of small oligomannose glycans, and Gal residues are capped by one or more Glc. In summary, E. histolytica N-glycans include unprocessed Man(5)GlcNAc(2), which is a target for cyanovirin-N, as well as unique, complex N-glycans containing Gal and Glc.
溶组织内阿米巴是一种可引发阿米巴痢疾和肝脓肿的原生生物,其N-聚糖因多种原因备受关注。溶组织内阿米巴能产生一种不同寻常的截短型N-聚糖前体(Man(5)GlcNAc(2)),核苷酸糖转运体数量稀少,其表面被凝集素伴刀豆球蛋白A所覆盖。在此,运用生化和质谱方法对溶组织内阿米巴的N-聚糖生物合成及最终的N-聚糖进行了研究,得出以下结论。未加工的Man(5)GlcNAc(2)是溶组织内阿米巴中含量最为丰富的N-聚糖,它通过N-聚糖特异性抗逆转录病毒凝集素蓝绿菌素-N在溶组织内阿米巴表面聚集成帽状。Glc(1)Man(5)GlcNAc(2)由UDP-葡萄糖:糖蛋白葡萄糖基转移酶合成,该酶是蛋白质折叠过程中保守的N-聚糖依赖性内质网质量控制系统的一部分,也存在于成熟的N-聚糖中。一种对苦马豆素敏感的α-甘露糖苷酶将一些N-聚糖修剪为双触角的Man(3)GlcNAc(2)。溶组织内阿米巴的复杂N-聚糖是通过在小寡甘露糖聚糖的双臂上添加α1,2-连接的半乳糖而形成的,半乳糖残基被一个或多个葡萄糖所覆盖。总之,溶组织内阿米巴的N-聚糖包括未加工的Man(5)GlcNAc(2)(它是蓝绿菌素-N的作用靶点)以及含有半乳糖和葡萄糖的独特复杂N-聚糖。