Using GFP--ligand fusions to measure receptor-mediated endocytosis in living cells.
Recombinant DNA technology has enabled the production of many types of chimeric proteins containing heterologous functional domains that have served a variety of useful capacities for cell biology research. Among proteins gaining wide use as a fusion partner is Aequorea victoria green fluorescent protein (GFP). GFP has been employed by numerous groups as a reporter gene for cell transfection and as an autofluorescent tag by recombinant fusion to foreign sequences. Here we describe the use of GFP as a tag for ligands, and provide examples of how purified recombinant GFP-ligand fusion proteins may be used to detect ligand-receptor interactions, including receptor-mediated endocytosis. Both its utility and limitations are discussed.
Medina-Kauwe
Lali K
LK
Department of Biochemistry, Institute for Genetic Medicine, University of Southern California Keck School of Medicine, Los Angeles, California 90033, USA.
Chen
Xinhua
X
D000818:Animals; D004705:Endocytosis; D049452:Green Fluorescent Proteins; D006801:Humans; D008024:Ligands; D008164:Luminescent Proteins / Q000235:genetics*; D016296:Mutagenesis; D011956:Receptors, Cell Surface / Q000502:physiology*; D011993:Recombinant Fusion Proteins
D008024:Ligands; D008164:Luminescent Proteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins