RRC ID 41917
Author Nagahama M, Hayashi S, Morimitsu S, Sakurai J.
Title Biological activities and pore formation of Clostridium perfringens beta toxin in HL 60 cells.
Journal 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.
Volume 278(38)
Pages 36934-41
Published 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
Times Cited 72
Human and Animal Cells P-815(RCB1167) HL60(RCB0041) Vero(RCB0001) CHO CHO-K1(RCB0285) COS-7(RCB0539) Intestine 407(RCB0553) HeLa(RCB0007) MDCK(RCB0995) PC-12(RCB0009)