Structures of the glycosylphosphatidylinositol membrane anchors from Aspergillus fumigatus membrane proteins
Structures of the glycosylphosphatidylinositol membrane anchors from Aspergillus fumigatus membrane proteins
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DOI：
10.1093/glycob/cwg004
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发表时间：
2003-03-01
期刊：
影响因子：
4.3
通讯作者：
Ferguson, MAJ
中科院分区：
文献类型：
--
作者：
Fontaine, T;Magnin, T;Ferguson, MAJ
糖基磷脂酰肌醇（GPI）锚定蛋白已被确定在所有的真核生物。在真菌中，GPIs的结构和生物合成研究仅限于酿酒酵母。在这篇文章中，四个GPI锚定蛋白纯化的膜制备的人类丝状真菌病原体烟曲霉。使用新的方法应用于蛋白质印迹蛋白条带，GPI结构表征ES-MS，荧光标记，HPLC，和特定的酶消化。A.烟曲霉GPI膜锚定物显示为主要含有植物鞘氨醇和单羟基化C-24：0脂肪酸的肌醇-磷酸神经酰胺。在酵母菌中，只有神经酰胺存在于A. Fumigatus的Gas 1 p同源物Gel 1 p也是如此。含有甘油二酯的酿酒酵母。助理烟曲霉GPI聚糖部分主要是与葡糖胺残基连接的线性五甘露糖结构：Manalpha 1 - 3 Manalpha 1 - 2 Manalpha 1 - 2 Manalpha 1 - 6 Manalpha 1 -4GlcN。
Glycosylphosphatidylinositol (GPI)-anchored proteins have been identified in all eukaryotes. In fungi, structural and biosynthetic studies of GPIs have been restricted to the yeast Saccharomyces cerevisiae. In this article, four GPI-anchored proteins were purified from a membrane preparation of the human filamentous fungal pathogen Aspergillus fumigatus. Using new methodology applied to western blot protein bands, the GPI structures were characterized by ES-MS, fluorescence labeling, HPLC, and specific enzymatic digestions. The phosphatidylinositol moiety of the A. fumigatus GPI membrane anchors was shown to be an inositol-phosphoceramide containing mainly phytosphingosine and monohydroxylated C-24:0 fatty acid. In constrast to yeast, only ceramide was found in the GPI anchor structures of A. fumigatus, even for Gel1p, a homolog of Gas1p in S. cerevisiae that contains diacylglycerol. The A. fumigatus GPI glycan moiety is mainly a linear pentomannose structure linked to a glucosamine residue: Manalpha1-3Manalpha1-2Manalpha1-2Manalpha1-6Manalpha1-4GlcN.