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.
Shams-Eldin
Hosam
H
Medizinisches Zentrum für Hygiene und Medizinische Mikrobiologie, Philipps-Universität Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, Germany.
Azzouz
Nahid
N
Niehus
Sebastian
S
Smith
Terry K
TK
Schwarz
Ralph T
RT
D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics; D017261:Glycosylphosphatidylinositols / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D018411:Spodoptera / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D017261:Glycosylphosphatidylinositols; D011994:Recombinant Proteins; D000581:Amidohydrolases; C087696:N-acetylglucosaminyl-phosphatidylinositol de-N-acetylase