Expression of Eukaryotic Membrane Proteins in Insect Cells.

Studying the structure and function of eukaryotic membrane proteins requires obtaining monodisperse, active proteins that can be used for further analysis. Eukaryotic expression systems are preferred because they provide the essential posttranslational modifications needed for proper folding and localization. Here, we describe effective approaches for the initial protein expression screening in insect cells of the human membrane protein PIGF, an adaptor protein involved in the biosynthesis of glycosylphosphatidylinositol anchors. The methodologies include construct design and cloning, followed by either transient transfection of insect cells or baculovirus preparation, protein expression, and detergent screening. Finally, the quality of the expressed protein is assessed by fluorescence size exclusion chromatography.