Clostridium botulinum type A haemagglutinin-positive progenitor toxin (HA(+)-PTX) binds to oligosaccharides containing Gal beta1-4GlcNAc through one subcomponent of haemagglutinin (HA1).
Haemagglutinin (HA) activity of Clostridium botulinum type A 19S and 16S toxins (HA-positive progenitor toxin; HA(+)-PTX) was characterized. HA titres against human erythrocytes of HA(+)-PTX were inhibited by the addition of lactose, D-galactose, N-acetyl-D-galactosamine and D-fucose to the reaction mixtures. A direct glycolipid binding test demonstrated that type A HA(+)-PTX strongly bound to paragloboside and some neutral glycolipids, but did not bind to gangliosides. Type A HA(+)-PTX also bound to asialoglycoproteins (asialofetuin, neuraminidase-treated transferrin), but not to sialoglycoproteins (fetuin, transferrin). Although glycopeptidase F treatment of asialofetuin abolished the binding of HA(+)-PTX, endo-alpha-N-acetylgalactosaminidase treatment did not. Thus these results can be interpreted as indicating that type A HA(+)-PTX detects and binds to Gal beta 1-4GlcNAc in paragloboside and the N-linked oligosaccharides of glycoproteins. Regardless of neuraminidase treatment, type A HA(+)-PTX bound to glycophorin A which is a major sialoglycoprotein on the surface of erythrocytes. Both native glycophorin A and neuraminidase-treated glycophorin A inhibited the binding of erythrocytes to type A HA(+)-PTX. Since the N:-linked oligosaccharide of glycophorin A is di-branched and more than 50% of this sugar chain is monosialylated, type A HA(+)-PTX probably bound to the unsialylated branch of the N-linked oligosaccharide of glycophorin A and agglutinated erythrocytes. One subcomponent of HA, designated HA1, did not agglutinate native erythrocytes, although it did bind to erythrocytes, paragloboside and asialoglycoproteins in a manner quite similar to that of HA(+)-PTX. These results indicate that type A HA(+)-PTX binds to oligosaccharides through HA1.
Inoue
Kaoru
K
Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan1.
Fujinaga
Yukako
Y
Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan1.
Honke
Koichi
K
Department of Biochemistry, Osaka University Medical School, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan2.
Arimitsu
Hideyuki
H
Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan1.
Mahmut
Nazira
N
Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan1.
Sakaguchi
Yoshihiko
Y
Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan1.
Ohyama
Tohru
T
Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri099-2422, Japan3.
Watanabe
Toshihiro
T
Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri099-2422, Japan3.
Inoue
Katsuhiro
K
Department of Food Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri099-2422, Japan3.
Oguma
Keiji
K
Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan1.
D000116:Acetylgalactosamine / Q000378:metabolism; D019274:Botulinum Toxins, Type A / Q000378:metabolism*; D002845:Chromatography; D003014:Clostridium botulinum / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005915:Globosides / Q000378:metabolism; D006017:Glycolipids / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006385:Hemagglutination Inhibition Tests; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D037102:Lectins; D009844:Oligosaccharides / Q000378:metabolism*; D011485:Protein Binding
D005915:Globosides; D006017:Glycolipids; D006023:Glycoproteins; D006388:Hemagglutinins; D037102:Lectins; D009844:Oligosaccharides; C112408:hemagglutinin I; C036363:paragloboside; D019274:Botulinum Toxins, Type A; D000116:Acetylgalactosamine