Characterization of a detergent-resistant surface lamina in cultured human fibroblasts

Treatment of cultured human fibroblasts with 0.5% Triton X-100 produces substratum-anchored cytoskeletal preparations consisting of cytoplasmic filaments, nucleus and a plasma membrane-derived surface lamina. The lamina was visualized in fluorescence microscopy with fluorochrome-coupled wheat germ agglutinin (WGA) as a lace-like structure, extending throughout the cell domain. It displayed a different organization at the ventral and dorsal surfaces of the cell, partially coaligning with bundles of actin and myosin filaments at the dorsal cell surface. At the ventral surface vinculin patches appeared to be included in the surface lamina. Polyacrylamide gel electrophoresis, combined with lectin reactivity studies and lectin affinity chromatography, revealed a 140 kD sialoglycoprotein as the major glycoprotein component of the surface lamina.