title: Visualizations of binding and internalization of two nonlinked protein components of botulinum C2 toxin in tissue culture cells.
abstract: Binding and internalization of two nonlinked components of botulinum C2 toxin were visualized in tissue culture cells with components directly labeled with fluorescence. The binding of both untrypsinized and trypsinized component II (UT-II and T-II, respectively) to common specific sites on the cell membrane was evidenced by competitive binding between fluorescence-labeled and unlabeled components. The distribution patterns of fluorescence-labeled T-II and UT-II after binding to cells at 37 degrees C were different; T-II clustered on the cell membrane and entered the cells in endosomes, whereas UT-II entered the cells inefficiently and not in vesicles and was distributed on the nuclear surface. The difference may be due to the multivalent property of T-II, which is not shared with UT-II. Fluorescence-labeled component I, which binds only to cells bound with T-II, entered cells by the same route as T-II did; both colocated on the same clusters on the cell membrane and also in the same vesicles in the cytoplasm. The present results suggest that component I of C2 toxin, which ADP-ribosylates cytoplasmic actin, directly binds to T-II but not to UT-II on the cell membrane and is internalized into cells together with T-II in the same endosomes.
authors.affiliation: Department of Veterinary Science, College of Agriculture, University of Osaka Prefecture, Japan.
authors.forename: I
authors.initials: I
authors.lastname: Ohishi
authors.forename: A
authors.initials: A
authors.lastname: Yanagimoto
mesh_terms: D000818:Animals; D001905:Botulinum Toxins / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D014709:Vero Cells
chemical_list: D005456:Fluorescent Dyes; D001905:Botulinum Toxins