Spatial organization at the cell surface.
Approaches are described for analysis of spatial organization of cell surface structure. Extraction of cells with nonionic and ionic detergents, chelating and chaotropic agents, salts, and reducing agents results in selective solubilization of surface proteins. Bisimidate disulfide-containing crosslinking reagents produce complexes containing surface proteins which can be analyzed by subsequent dissociation of the complexes. Disulfide-bonded complexes are also found without addition of crosslinkers, and reducing agents aid in extracting surface proteins. These results suggest a possible role for disulfide bonds in cell surface organization. Immunofluorescent staining of cells with antisera to LETS protein and to actin reveals fibrillar structures which survive NP40 extraction. These results indicate a complex organization at the cell surface which is amenable to analysis by permutations of the methods described.
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure*; D002471:Cell Transformation, Neoplastic; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000032:analysis; D005455:Fluorescent Antibody Technique; D008565:Membrane Proteins / Q000032:analysis*; D008970:Molecular Weight; D009858:Oncogenic Viruses
D008565:Membrane Proteins