An efficient method to express GPI-anchor proteins in insect cells

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• 作者：Hosam Shams-Eldin ; Nahid Azzouz ; Sebastian Niehus ; Terry K. Smith ; Ralph T. Schwarz
• 关键词：Baculovirus ; Insect cells ; Sf9 ; High Five ; Glycosylphosphatidylinositols ; N-glycosylation ; Toxoplasma gondii ; N-acetylglucosaminyl phosphatidylinositol de-N-acetylase ; PIGL ; SAG1
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2008
• 出版时间：25 January 2008
• 年：2008
• 卷：365
• 期：4
• 页码：657-663
• 全文大小：500 K

文摘

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.