Mobility and distribution of a cell surface glycoprotein and its interaction with other membrane components

Abstract

Fluorescence photobleaching recovery and immunofluorescence methods were used to study the lateral mobility and topographical distribution of a major cell surface glycoprotein (CSP) [chick embryo fibroblast CEF cells, Swiss mouse fibroblast 3T3 cells]. Endogenous CSP and fluorescent-labeled exogenous CSP bind to the cell surface in a fibrillar pattern and were immobile on the experimental time scale. Azide, vinblastine and cytochalasin B do not alter the immobility and cell surface distribution of the CSP molecules. Oxidative phosphorylation and the cytoskeleton do not seem to be responsible for the properties of the bound glycoprotein. The presence of immobile CSP fibrils does not impede the diffusion of a lipid probe, a ganglioside analogue or various surface antigens. The fibrils apparently do not form a barrier across the lipid phase of the plasma membrane. Concanavalin A binds to CSP and is largely immobile in regions rich in CSP. The presence of immobile concanavalin A receptors in areas or on cells lacking CSP indicates that other types of immobile concanavalin A receptors also exist. CSP does not bind to lipid bilayers composed of phosphatidylcholine or oxidized cholesterol. It does bind to dextran-coated bilayers as a diffuse distribution of mobile molecules that can patch after addition of antibodies to CSP. CSP molecules probably do not interact strongly with other CSP molecules under these conditions. Exogenous CSP binds to regions on the cell surface that already bear CSP. In view of the apparent weakness of CSP-CSP interactions on the lipid bilayer, it seems possible that the assembly of CSP fibrils is nucleated by cell surface components in addition to CSP.