Studies on the in vitro uncoating of poliovirus. IV. Characteristics of solubilized membrane-modifying and -stabilizing factors.

Previous studies in our laboratory on the in vitro uncoating of poliovirus have shown that HeLa cell membrane contains a modifying factor which induces early modification of virus (the loss of VP4) and a stabilizing factor which protects virus against heat-induced degradation. It has now been found that membrane-modifying factor is dependent on the presence of phospholipid for activity. Modifying activity was lost after exposure of membrane (Mem) to phospholipase C. Triton X-100-solubilized modifying factor prepared from phospholipase C-treated Mem was reactivated by phospholipid. Lecithin, phosphatidylethanolamine, phosphatidylserine, and sphingomyelin were found to exhibit a reactivating effect. Lecithin was the most effective individual phospholipid in terms of maximal reactivation. Partial purification of Triton X-100-solubilized modifying factor was achieved by concanavalin A-Sepharose chromatography. Membrane-stabilizing factor was extracted from HeLa cell membrane by solubilization with sodium deoxycholate (DOC). Some properties of DOC-solubilized stabilizing factor were studied. The solubilized stabilizing factor was inactivated by treatment with trypsin or chymotrypsin. Treatment of the solubilized stabilizing factor with certain lipid solvents, lipolytic enzymes, or lectins had no detectable effect on stabilizing activity.