The use of a fluorescent lipid as a non-covalent bound tracer of integral membrane proteins.
Solubilized and affinity purified major histocompatibility class I and class II antigens (MHC-I, MHC-II) were incorporated into liposomes, which contained a fluorescent lipid, by adsorption of the detergent to Biobeads SM-2. The fluorescent lipid was used as a non-covalently bound tracer to detect antibody-mediated agglutination of liposomes with MHC antigens. After incorporation of the membrane proteins into liposomes of different lipid composition, phosphatidylcholine was chosen as a lipid matrix. The use of a fluorescent lipid as a tracer of membrane proteins was demonstrated by the detection of human beta 2-microglobulin association with mouse MHC-I. Furthermore the method was used to measure the amount of liposomes containing two different membrane proteins.
Eriksson
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Cancer Biology Laboratory, State University Hospital (Rigshospitalet), Copenhagen, Denmark.
D011232:Chemical Precipitation; D005456:Fluorescent Dyes; D006183:H-2 Antigens / Q000378:metabolism; D008055:Lipids; D008081:Liposomes; D008565:Membrane Proteins / Q000032:analysis*; D011485:Protein Binding; D012995:Solubility; D001613:beta 2-Microglobulin / Q000378:metabolism
D005456:Fluorescent Dyes; D006183:H-2 Antigens; D008055:Lipids; D008081:Liposomes; D008565:Membrane Proteins; D001613:beta 2-Microglobulin