Characterization of Vibrio cholerae El Tor cytolysin as an oligomerizing pore-forming toxin.
V. cholerae El Tor cytolysin is a secreted, water-soluble protein of M(r) 60,000 that may be relevant to the pathogenesis of acute diarrhea. In this communication, we demonstrate that the toxin binds to and oligomerizes in target membranes to form SDS-stable aggregates of M(r) 200,000-250,000 that generate small transmembrane pores. Pores formed in erythrocytes were approximately 0.7 nm in size, as demonstrated by osmotic protection experiments. Binding was shown to occur in a temperature-independent manner preceding the temperature-dependent oligomerization step. Pores were also shown to be formed in L929 and HEp-2 cells, human fibroblasts and keratinocytes, albeit with highly varying efficacy. At neutral pH and in the presence of serum, human fibroblasts were able to repair a limited number of lesions. The collective data identify V. cholerae El Tor cytolysin as an oligomerizing toxin that damages cells by creating small transmembrane pores.
Zitzer
A
A
Institute of Medical Microbiology and Hygiene, Mainz, Germany.
Walev
I
I
Palmer
M
M
Bhakdi
S
S
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D003603:Cytotoxins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006461:Hemolysis; D006801:Humans; D007473:Ion Channels; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011108:Polymers; D011817:Rabbits; D012967:Sodium Dodecyl Sulfate; D014734:Vibrio cholerae / Q000737:chemistry*
D003603:Cytotoxins; D007473:Ion Channels; D011108:Polymers; D012967:Sodium Dodecyl Sulfate; D000255:Adenosine Triphosphate