FIGURE 2.

Binding of aldolase, GAPDH, LDH, PK, and ovalbumin to fresh porous ghosts. Sulfo-SBED-labeled aldolase, GAPDH, LDH, PK, and ovalbumin were incubated with fresh ghosts, as described under “Experimental Procedures,” and after washing and photoactivating, bound proteins were separated by SDS-PAGE and stained with Coomassie Blue (A) or transferred to nitrocellulose and incubated with streptavidin-HRP to reveal biotinylated proteins (B). Because ovalbumin does not bind to erythrocyte membranes, washing to remove unbound protein prevents labeling of any membrane proteins with sulfo-SBED-ovalbumin. Lane 1, molecular weight markers; lanes 2–6, ghosts that were incubated with sulfo-SBED-aldolase (lane 2), sulfo-SBED-GAPDH (lane 3), sulfo-SBED-LDH (lane 4), sulfo-SBED-PK (lane 5), and sulfo-SBED-ovalbumin (lane 6).