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An efficient method to express GPI-anchor proteins in insect cells |
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| Authors: | Shams-Eldin Hosam Azzouz Nahid Niehus Sebastian Smith Terry K Schwarz Ralph T |
| Institution: | a Medizinisches Zentrum für Hygiene und Medizinische Mikrobiologie, Philipps-Universität Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, Germany b Laboratory for Organic Chemistry, Swiss Federal Institute of Technology ETH Zürich, Wolfgang-Pauli-Strasse 10, CH-8093 Zürich, Switzerland c Center for Biomolecular Sciences, The North Haugh, The University, St. Andrews. KY16 9ST, Scotland, UK d Unité de Glycobiologie Structurale et Fonctionnelle UMR CNRS/USTL n° 8576-IFR 147, Université des Sciences et Technologies de Lille 59655, Villeneuve D’Ascq Cedex, France |
| Abstract: | 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. |
| Keywords: | Baculovirus Insect cells Sf9 High Five Glycosylphosphatidylinositols N-glycosylation Toxoplasma gondii N-acetylglucosaminyl phosphatidylinositol de-N-acetylase PIGL SAG1 |
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