Detergent-resistant membranes are platforms for actinoporin pore-forming activity on intact cells

被引:46
作者
Alegre-Cebollada, J [1 ]
Rodríguez-Crespo, I [1 ]
Gavilanes, JG [1 ]
del Pozo, AM [1 ]
机构
[1] Univ Complutense Madrid, Fac Quim, Dept Bioquim, E-28040 Madrid, Spain
关键词
COS-7; cells; cyclodextrin; equinatoxin II; Stichodactyla helianthus; sticholysin II;
D O I
10.1111/j.1742-4658.2006.05122.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Sticholysin II is a pore-forming toxin produced by the sea anemone Stichodactyla helianthus. We studied its cytolytic activity on COS-7 cells. Fluorescence spectroscopy and flow cytometry revealed that the toxin permeabilizes cells to propidium cations in a dose-dependent and time-dependent manner. This permeabilization is impaired by preincubation of cells with cyclodextrin. Isolation of detergent-resistant cellular membranes showed that sticholysin II colocalizes with caveolin-1 in fractions corresponding to raft-like domains. The interaction of sticholysin II with such domains is only lipid dependent as it also occurs in the absence of any other membrane-associated protein. Toxin binding to raft-like lipid vesicles inhibited cell permeabilization. The results suggest that sticholysin II promotes pore formation in COS-7 cells through interaction with membrane domains which behave like cellular rafts.
引用
收藏
页码:863 / 871
页数:9
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