title: Cross-reactivity of anthrax and C2 toxin: protective antigen promotes the uptake of botulinum C2I toxin into human endothelial cells.
abstract: Binary toxins are among the most potent bacterial protein toxins performing a cooperative mode of translocation and exhibit fatal enzymatic activities in eukaryotic cells. Anthrax and C2 toxin are the most prominent examples for the AB(7/8) type of toxins. The B subunits bind both host cell receptors and the enzymatic A polypeptides to trigger their internalization and translocation into the host cell cytosol. C2 toxin is composed of an actin ADP-ribosyltransferase (C2I) and C2II binding subunits. Anthrax toxin is composed of adenylate cyclase (EF) and MAPKK protease (LF) enzymatic components associated to protective antigen (PA) binding subunit. The binding and translocation components anthrax protective antigen (PA(63)) and C2II of C2 toxin share a sequence homology of about 35%, suggesting that they might substitute for each other. Here we show by conducting in vitro measurements that PA(63) binds C2I and that C2II can bind both EF and LF. Anthrax edema factor (EF) and lethal factor (LF) have higher affinities to bind to channels formed by C2II than C2 toxin's C2I binds to anthrax protective antigen (PA(63)). Furthermore, we could demonstrate that PA in high concentration has the ability to transport the enzymatic moiety C2I into target cells, causing actin modification and cell rounding. In contrast, C2II does not show significant capacity to promote cell intoxication by EF and LF. Together, our data unveiled the remarkable flexibility of PA in promoting C2I heterologous polypeptide translocation into cells.
authors.affiliation: Rudolf-Virchow-Center, University of Würzburg, Würzburg, Germany.
authors.forename: Angelika
authors.initials: A
authors.lastname: Kronhardt
authors.forename: Monica
authors.initials: M
authors.lastname: Rolando
authors.forename: Christoph
authors.initials: C
authors.lastname: Beitzinger
authors.forename: Caroline
authors.initials: C
authors.lastname: Stefani
authors.forename: Michael
authors.initials: M
authors.lastname: Leuber
authors.forename: Gilles
authors.initials: G
authors.lastname: Flatau
authors.forename: Michel R
authors.initials: MR
authors.lastname: Popoff
authors.forename: Roland
authors.initials: R
authors.lastname: Benz
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Lemichez
mesh_terms: D036002:ADP Ribose Transferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D001905:Botulinum Toxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; C030325:anthrax toxin; D036002:ADP Ribose Transferases; D001905:Botulinum Toxins; D000262:Adenylyl Cyclases