Staphylococcal alpha-toxin kills human keratinocytes by permeabilizing the plasma membrane for monovalent ions.
Incubation of human keratinocytes with nanomolar concentrations of Staphylococcus aureus alpha-toxin leads to irreversible depletion of cellular ATP. The toxin forms hexamers in the target cell membranes, and rapid transmembrane flux of K+, Na+, and 86Rb+ is observed. Unexpectedly, pores formed in keratinocytes through application of low but lethal doses of alpha-toxin appeared to be considerably smaller than those formed in erythrocyte membranes. They permitted neither rapid influx of Ca2+ or propidium iodide, nor efflux of carboxyfluorescein. Larger pores allowing flux of all three markers did form when the toxin was applied at high concentrations. Flux of monovalent ions and reduction in cellular ATP levels evoked by low toxin doses correlated temporally with a fall in oxygen consumption, which was interpreted to reflect breakdown of mitochondrial respiration. The lethal event could not be thwarted by manipulating the extracellular K+ or Ca2+ concentrations. Realization that alpha-toxin may form very small pores in nucleated cells is important for future research on cellular toxin effects and membrane repair processes.
Walev
I
I
Institute of Medical Microbiology, University of Mainz, Germany.
Martin
E
E
Jonas
D
D
Mohamadzadeh
M
M
Müller-Klieser
W
W
Kunz
L
L
Bhakdi
S
S
D001427:Bacterial Toxins / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D002118:Calcium / Q000378:metabolism; D002414:Cations, Monovalent / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D003470:Culture Media; D006460:Hemolysin Proteins / Q000737:chemistry / Q000633:toxicity*; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D007700:Kinetics; D008968:Molecular Conformation; D008970:Molecular Weight; D010101:Oxygen Consumption / Q000187:drug effects; D011188:Potassium / Q000378:metabolism; D013211:Staphylococcus aureus
D001427:Bacterial Toxins; D002414:Cations, Monovalent; D003470:Culture Media; D006460:Hemolysin Proteins; C021091:staphylococcal alpha-toxin; D011188:Potassium; D002118:Calcium