| RRC ID |
43825
|
| 著者 |
Miyata K, Sagane Y, Inui K, Miyashita S, Suzuki T, Oguma K, Ohyama T, Niwa K, Watanabe T.
|
| タイトル |
Purification and characterization of nontoxic protein complex from serotype D 4947 botulinum toxin complex.
|
| ジャーナル |
Protein J
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| Abstract |
The large-sized botulinum toxin complex (L-TC) is composed of botulinum neurotoxin (BoNT) and nontoxic proteins, e.g. nontoxic nonhemagglutinin (NTNHA) and three types of hemagglutinins (HAs; HA-33, HA-17 and HA-70). The nontoxic proteins play a critical role in L-TC oral toxicity by protecting the BoNT in the digestive tract, and facilitating absorption of the L-TC across the intestinal wall. Under alkaline conditions, the L-TC separates into BoNT and the nontoxic protein complex (NC). In this study, we established a two-step procedure to yield highly pure NC from the L-TC produced by Clostridium botulinum serotype D strain 4947 in which the NC was isolated from the L-TC by gel filtration under alkaline conditions followed by immunoprecipitation with an anti-BoNT antibody to remove contaminating BoNT from the NC fraction. Western blotting and electrophoretic analysis showed that the highly purified NC fraction had only very slight or no BoNT contamination. In addition, the purified NC fraction showed no intraperitoneal (ip) toxicity to mice at a dose of 38 ng per animal whereas the L-TC exhibited an ip median lethal dose of 0.38 ng per mouse. The highly purified NC displayed the same hemagglutination titer as the L-TC. The NC, as well as the L-TC, demonstrated cell binding and monolayer transport in the rat intestinal epithelial cell line IEC-6. Consequently, the highly purified NC can function as a "delivery vehicle" even without the BoNT.
|
| 巻・号 |
31(5)
|
| ページ |
387-92
|
| 公開日 |
2012-6-1
|
| DOI |
10.1007/s10930-012-9413-x
|
| PMID |
22565346
|
| MeSH |
Animals
Botulinum Toxins / chemistry*
Botulinum Toxins / isolation & purification
Botulinum Toxins / metabolism
Botulinum Toxins / toxicity*
Botulism / microbiology
Cell Line
Chromatography, Gel
Clostridium botulinum type D / chemistry*
Clostridium botulinum type D / metabolism
Electrophoresis, Polyacrylamide Gel
Hemagglutination / drug effects
Hemagglutination Tests
Horses
Immunoprecipitation
Mice
Mice, Inbred BALB C
Rats
|
| IF |
1.317
|
| 引用数 |
3
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
|
