Binding properties of Clostridium botulinum type C progenitor toxin to mucins.
It has been reported that Clostridium botulinum type C 16S progenitor toxin (C16S toxin) first binds to the sialic acid on the cell surface of mucin before invading cells [A. Nishikawa, N. Uotsu, H. Arimitsu, J.C. Lee, Y. Miura, Y. Fujinaga, H. Nakada, T. Watanabe, T. Ohyama, Y. Sakano, K. Oguma, The receptor and transporter for internalization of Clostridium botulinum type C progenitor toxin into HT-29 cells, Biochem. Biophys. Res. Commun. 319 (2004) 327-333]. In this study we investigated the binding properties of the C16S toxin to glycoproteins. Although the toxin bound to membrane blotted mucin derived from the bovine submaxillary gland (BSM), which contains a lot of sialyl oligosaccharides, it did not bind to neuraminidase-treated BSM. The binding of the toxin to BSM was inhibited by N-acetylneuraminic acid, N-glycolylneuraminic acid, and sialyl oligosaccharides strongly, but was not inhibited by neutral oligosaccharides. Both sialyl alpha2-3 lactose and sialyl alpha2-6 lactose prevented binding similarly. On the other hand, the toxin also bound well to porcine gastric mucin. In this case, neutral oligosaccharides might play an important role as ligand, since galactose and lactose inhibited binding. These results suggest that the toxin is capable of recognizing a wide variety of oligosaccharide structures.
Nakamura
Toshio
T
Department of Applied Biological Science and Department of Biotechnology, United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi, Tokyo 183-8509, Japan.
Takada
Noriko
N
Tonozuka
Takashi
T
Sakano
Yoshiyuki
Y
Oguma
Keiji
K
Nishikawa
Atsushi
A
D000818:Animals; D001667:Binding, Competitive; D001905:Botulinum Toxins / Q000737:chemistry* / Q000378:metabolism; D002236:Carbohydrate Conformation; D002417:Cattle; D005752:Gastric Mucins / Q000737:chemistry; D009077:Mucins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D009438:Neuraminic Acids / Q000737:chemistry; D009439:Neuraminidase / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry; D011485:Protein Binding; D013363:Submandibular Gland / Q000737:chemistry; D013552:Swine; D014168:Transferrin / Q000737:chemistry
D005752:Gastric Mucins; D009077:Mucins; D009438:Neuraminic Acids; D009844:Oligosaccharides; D014168:Transferrin; C016735:sialooligosaccharides; C032592:N-glycolylneuraminic acid; D009439:Neuraminidase; D001905:Botulinum Toxins; C096324:botulinum toxin type C; D019158:N-Acetylneuraminic Acid