Cytotoxicity of Clostridium septicum alpha-toxin: its oligomerization in detergent resistant membranes of mammalian cells

Alpha-toxin is an important agent of the virulence of Clostridium septicum. We examined cytotoxicity for alpha-toxin to various mammalian cells with recombinant toxin fused with a histidine-tag at the amino-terminal. The recombinant toxin retained the activity indistinguishable from the native form....

Full description

Saved in:
Bibliographic Details
Published in:Microbial pathogenesis 2004-12, Vol.37 (6), p.279-286
Main Authors: Hang'ombe, Mudenda B., Mukamoto, Masafumi, Kohda, Tomoko, Sugimoto, Nakaba, Kozaki, Shunji
Format: Article
Language:English
Subjects:
Alpha-toxin
animal pathogenic bacteria
Animals
bacterial toxins
Bacterial Toxins - genetics
Bacterial Toxins - metabolism
Bacterial Toxins - toxicity
beta-Cyclodextrins - pharmacology
Biological and medical sciences
Cattle
Cell Line
Cell Membrane - drug effects
Cell Membrane - metabolism
Clostridium septicum
Cricetinae
Cytotoxicity
Detergent-resistant membranes
Detergents - pharmacology
Dimerization
Dogs
Fundamental and applied biological sciences. Psychology
Mice
Microbiology
Oligomerization
Rats
Recombinant Proteins - metabolism
Recombinant Proteins - toxicity
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Alpha-toxin is an important agent of the virulence of Clostridium septicum. We examined cytotoxicity for alpha-toxin to various mammalian cells with recombinant toxin fused with a histidine-tag at the amino-terminal. The recombinant toxin retained the activity indistinguishable from the native form. Mammalian nucleated cells examined in this study are more sensitive to the protoxin than to the trypsinized toxin, except RAW 264.7 and P3U1 cells of myeloid lineage. Cellular proteins of various molecular sizes interacted with the toxin. The size and SDS-PAGE pattern of the proteins were different among cell lines but they were liberated from the cells by the treatment with phosphatidylinositol-specific phospholipase C. The toxin appeared to target and utilize detergent resistant membranes (DRMs) for binding and subsequent oligomerization. In discontinuous sucrose density gradient, we demonstrated by immunoblotting that the toxin bound to DRMs contained in L929 cells and caused the oligomer formation. Furthermore, cholesterol depletion with cholesterol-interacting agents reduced toxin oligomerization and lowered cytotoxicity of the toxin towards cells. These results suggest that alpha-toxin preferentially exploits DRMs for oligomerization.
ISSN:0882-4010
1096-1208
DOI:10.1016/j.micpath.2004.09.001