Incorporation of Artificial Lipid-Anchored Proteins into Cultured Mammalian Cells

Exogenous lipid-anchored proteins can be incorporated into the plasma membranes of living mammalian cells, allowing the chemical structure of the incorporated protein and its lipid anchor to be controlled (and varied) to a much greater degree than is possible for proteins expressed by the cells themselves. This technology offers a variety of potential applications, including incorporating novel and complex protein constructs into cell surfaces and exploring structure-function relationships for biologically important lipid-anchored proteins such as glycosylphosphatidylinositol-anchored proteins. Here we describe detailed methods for stable incorporation of artificial lipid-anchored proteins into cultured mammalian cells under mild, nonperturbing conditions.