| RRC ID |
41917
|
| 著者 |
Nagahama M, Hayashi S, Morimitsu S, Sakurai J.
|
| タイトル |
Biological activities and pore formation of Clostridium perfringens beta toxin in HL 60 cells.
|
| ジャーナル |
J Biol Chem
|
| Abstract |
Clostridium perfringens beta toxin is an important agent of necrotic enteritis. Of the 10 cell lines tested, only the HL 60 cell line was susceptible to beta toxin. The toxin induced swelling and lysis of the cell. Treatment of the cells with the toxin resulted in K+ efflux from the cells and Ca2+, Na+, and Cl- influxes. These events reached a maximum just before the cells were lysed by the toxin. Incubation of the cells with the toxin showed the formation of toxin complexes of about 191 and 228 kDa, which were localized in the domains that fulfilled the criteria of lipid rafts. The complex of 228 kDa was observed until 30 min after incubation, and only the complex of 191 kDa was remained after 60 min. Treatment of the cells with methyl-beta-cyclodextrin or cholesterol oxidase blocked binding of the toxin to the rafts and the toxin-induced K+ efflux and swelling. The toxin-induced Ca2+ influx and morphological changes were inhibited by an increase in the hydrodynamic diameter of polyethylene glycols from 200 to 400 and markedly or completely inhibited by polyethylene glycol 600 and 1000. However, these polyethylene glycols had no effect on the toxin-induced K+ efflux. The toxin induced carboxyfluorescein release from phosphatidyl-choline-cholesterol liposomes containing carboxyfluorescein and formed an oligomer with 228 kDa in a dose-dependent manner but did not form an oligomer with the 191-kDa complex. We conclude that the toxin acts on HL 60 cells by binding to lipid rafts and forming a functional oligomer with 228 kDa.
|
| 巻・号 |
278(38)
|
| ページ |
36934-41
|
| 公開日 |
2003-9-19
|
| DOI |
10.1074/jbc.M306562200
|
| PII |
S0021-9258(20)83511-X
|
| PMID |
12851396
|
| MeSH |
Amino Acid Sequence
Animals
Bacterial Toxins / chemistry*
Bacterial Toxins / pharmacology*
CHO Cells
COS Cells
Calcium / chemistry
Calcium / metabolism
Cell Line
Cell Line, Tumor
Centrifugation, Density Gradient
Chlorine / metabolism
Chlorocebus aethiops
Cholesterol / metabolism
Cricetinae
Dogs
Dose-Response Relationship, Drug
HL-60 Cells
HeLa Cells
Humans
Immunoblotting
Ions
L-Lactate Dehydrogenase / metabolism
Liposomes / metabolism
Membrane Microdomains / metabolism
Mice
Molecular Sequence Data
PC12 Cells
Phosphatidylcholines / metabolism
Polyethylene Glycols / chemistry
Potassium / chemistry
Potassium / metabolism
Protein Binding
Protein Structure, Tertiary
Rats
Sucrose / pharmacology
Time Factors
Vero Cells
|
| IF |
4.238
|
| 引用数 |
72
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
P-815(RCB1167)
HL60(RCB0041)
Vero(RCB0001)
CHO
CHO-K1(RCB0285)
COS-7(RCB0539)
Intestine 407(RCB0553)
HeLa(RCB0007)
MDCK(RCB0995)
PC-12(RCB0009) |
