ABSTRACT
Amantadine and related compounds stabilized the structure of purified pig brain clathrin coated vesicles (CCV) at biologically relevant concentrations. Incubation of purified CCV for 30 min at 25 degrees C or 37 degrees C caused the release of clathrin, as determined by a centrifugation assay, and a reduction in the number of coated vesicles, by electron microscopy. Amantadine (10 mM), tromantadine (1 mM), amidine D295 (cyclohexylcarboximidamide-(N-benzyl)hydrochloride (10 mM), chloroquine (0.1 mM) and monodansylcadaverine (10 mM) significantly reduced the extent of dissociation.
Subject(s)
Amantadine/analogs & derivatives , Amantadine/pharmacology , Amines/pharmacology , Clathrin/metabolism , Coated Pits, Cell-Membrane/metabolism , Animals , Brain/metabolism , Coated Pits, Cell-Membrane/drug effects , Kinetics , Structure-Activity Relationship , SwineABSTRACT
Addition of tromantadine after virus penetration inhibited HSV-1 induced syncytium formation and virus production in HEp-2 and VERO cells and acted additively with neutralizing antibody in blocking virus spread and cytopathology. Inhibition of syncytium formation in VERO cells infected with 0.01 pfu/cell of HSV-1 GC+ was observed at a concentration greater than 25 micrograms/ml. The extent of inhibition was dependent upon the multiplicity of infection and cell type. Tromantadine inhibited a late event in HSV-1 replication which appeared to be sensitive to cycloheximide. Reversal of the inhibitory effect of tromantadine on syncytium formation required new protein synthesis. HSV-1 gB, gC, and gD were synthesized in the presence of tromantadine and could be detected on the cell surface by immunofluorescence. Tromantadine most likely inhibits a cellular process that is required for syncytium formation, such as glycoprotein processing, which occurs after the synthesis of the fusion protein but before its expression on the cell surface.
