H-2Kk and vesicular stomatitis virus G proteins are not extensively associated in reconstituted membranes recognized by T cells.
It is shown that liposomes containing (i) a fluorescein-labeled murine histocompatibility antigen (FITC-H-2Kk) and the G protein of vesicular stomatitis virus or (ii) H-2Kk and fluorescein-labeled viral protein (FITC-G) can elicit H-2-restricted syngeneic antiviral cytotoxic T cells as assayed by 51Cr release from appropriate virus-infected target cells. Fluorescence recovery after photobleaching was used to measure the diffusion coefficients of these reconstituted proteins in four different samples: (i) FITC-H-2Kk; (ii) FITC-H-2Kk and G; (iii) FITC-G; and (iv) FITC-G and H-2Kk. The same rate of lateral diffusion (D = 1 x 10(-8) cm2/sec at 37 degrees C in 25% cholesterol/75% dimyristoylphosphatidylcholine) was obtained in every case. Both proteins, fluorescent as well as nonfluorescent, could be patched by using specific antibodies. When G was patched with antibody, FITC-H-2Kk did not copatch. When H-2Kk was patched with antibody FITC-G did not copatch. These diffusion and patching measurements rule out the possibility that these proteins have either extensive oligomeric associations or strong specific pairwise associations.
D000818:Animals; D000937:Antigen-Antibody Reactions; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure*; D004058:Diffusion; D006183:H-2 Antigens / Q000276:immunology / Q000378:metabolism*; D008081:Liposomes; D008560:Membrane Fluidity; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D013601:T-Lymphocytes / Q000276:immunology; D014721:Vesicular stomatitis Indiana virus; D014764:Viral Proteins / Q000276:immunology / Q000378:metabolism*
D006183:H-2 Antigens; D008081:Liposomes; D008565:Membrane Proteins; D014764:Viral Proteins