Shams-Eldin Hosam, Azzouz Nahid, Niehus Sebastian, Smith Terry K, Schwarz Ralph T
Medizinisches Zentrum für Hygiene und Medizinische Mikrobiologie, Philipps-Universität Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, Germany.
Biochem Biophys Res Commun. 2008 Jan 25;365(4):657-63. doi: 10.1016/j.bbrc.2007.11.026. Epub 2007 Nov 20.
Glycosylphosphatidylinositols (GPIs) constitute a class of glycolipids that have various functions, the most basic being to attach proteins to the surface of eukaryotic cells. GPIs have to be taken into account, when expressing surface antigens from parasitic protozoa in heterologous systems. The synthesis of the GPI-anchors was previously reported to be drastically decreased to almost background level following baculovirus infection. Here we describe a new method to express GPI-anchor proteins in insect cells relying on using of a supplementary baculovirus construct that overexpresses the N-acetylglucosaminyl phosphatidylinositol de-N-acetylase, the enzyme catalyzing the second step in the GPI biosynthetic pathway.
糖基磷脂酰肌醇(GPI)构成一类具有多种功能的糖脂,其最基本的功能是将蛋白质附着于真核细胞表面。在异源系统中表达寄生原生动物的表面抗原时,必须考虑GPI。先前有报道称,杆状病毒感染后,GPI锚的合成急剧减少至几乎背景水平。在此,我们描述了一种在昆虫细胞中表达GPI锚蛋白的新方法,该方法依赖于使用一种补充杆状病毒构建体,该构建体过表达N-乙酰葡糖胺基磷脂酰肌醇脱N-乙酰酶,该酶催化GPI生物合成途径中的第二步。