Characterization of toxin complex produced by a unique strain of Clostridium botulinum serotype D 4947.
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.
Hasegawa
Kimiko
K
Department of Food Science and Technology, Faculty of Bioindustry, Tokyo University of Agriculture, Abashiri 099-2493, Japan.
Watanabe
Toshihiro
T
Sato
Hiroaki
H
Sagane
Yoshimasa
Y
Mutoh
Shingo
S
Suzuki
Tomonori
T
Yamano
Akihito
A
Kouguchi
Hirokazu
H
Takeshi
Kouichi
K
Kamaguchi
Arihide
A
Fujinaga
Yukako
Y
Oguma
Keiji
K
Ohyama
Tohru
T
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
D001427:Bacterial Toxins; D006388:Hemagglutinins; D009498:Neurotoxins; D011506:Proteins; D014357:Trypsin; D001905:Botulinum Toxins