Biochemical and Biophysical Research Communications, Vol.365, No.4, 657-663, 2008
An efficient method to express GPI-anchor proteins in insect cells
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. (c) 2007 Elsevier Inc. All rights reserved.
Keywords:baculovirus;insect cells;Sf9;high five;glycosylphosphatidylinositols;N-glycosylation;toxoplasma gondii;N-acetylglucosaminyl phosphatidylinositol de-n-acetylase;PIGL;SAG1