title: Characterization of toxin complex produced by a unique strain of Clostridium botulinum serotype D 4947.
abstract: A unique strain of Clostridium botulinum, serotype D 4947 (D-4947), produces a considerable amount of a 650 kDa toxin complex (L-TC) and a small amount of a 280 kDa M-TC, a 540 kDa TC, and a 610 kDa TC. The complexes are composed of only un-nicked components, including neurotoxin (NT), nontoxic nonhemagglutinin (NTNHA) and hemagglutinin subcomponents (HA-70, HA-33 and HA-17). Unlike other NTs from all serotype strains, separation of D-4947 NT from L-TC, except for M-TC, during chromatography required highly alkaline conditions around pH 8.8. The separated NT and NTNHA/HAs complex can be reconstituted to L-TC that is indistinguishable from the parent L-TC with respect to toxicity, hemagglutination activity and gel filtration profile. The isoelectric points of NT and NTNHA/HAs were close together depending on the number of HA-33/17 molecules. We have established a new method to separate the unique D-4947 NT from the complex, which will yield valuable information on structure of botulinum toxin.
authors.affiliation: Department of Food Science and Technology, Faculty of Bioindustry, Tokyo University of Agriculture, Abashiri 099-2493, Japan.
authors.forename: Kimiko
authors.initials: K
authors.lastname: Hasegawa
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Watanabe
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Sato
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Sagane
authors.forename: Shingo
authors.initials: S
authors.lastname: Mutoh
authors.forename: Tomonori
authors.initials: T
authors.lastname: Suzuki
authors.forename: Akihito
authors.initials: A
authors.lastname: Yamano
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kouguchi
authors.forename: Kouichi
authors.initials: K
authors.lastname: Takeshi
authors.forename: Arihide
authors.initials: A
authors.lastname: Kamaguchi
authors.forename: Yukako
authors.initials: Y
authors.lastname: Fujinaga
authors.forename: Keiji
authors.initials: K
authors.lastname: Oguma
authors.forename: Tohru
authors.initials: T
authors.lastname: Ohyama
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000737:chemistry*; D001905:Botulinum Toxins / Q000737:chemistry* / Q000302:isolation & purification; D002850:Chromatography, Gel; D003014:Clostridium botulinum / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006388:Hemagglutinins / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D051379:Mice; D009498:Neurotoxins / Q000737:chemistry; D010537:Peritoneum / Q000187:drug effects; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry; D013045:Species Specificity; D014357:Trypsin / Q000737:chemistry
chemical_list: D001427:Bacterial Toxins; D006388:Hemagglutinins; D009498:Neurotoxins; D011506:Proteins; D014357:Trypsin; D001905:Botulinum Toxins