Oligomerization of 3H-labelled staphylococcal alpha-toxin and fragments on adrenocortical Y1 tumour cells.
Staphylococcus aureus alpha-toxin has previously been shown to bind to erythrocyte membranes and the isolated membranes contain the toxin in both monomeric and hexameric form. The hexamers are believed to form the ring-shaped structures observed by electron microscopy on toxin-treated erythrocytes. It has not previously been shown that hexamers are formed also on nucleated mammalian cells although it has been assumed that hexamers in both systems create transmembrane channels, responsible for the toxin-induced membrane damage. Here we demonstrate by autoradiography that 3H-alpha-toxin bound to and formed high molecular weight complexes-presumably hexamers-on cultured adrenocortical Y1 tumour cells. The binding kinetics suggested a non-specific association of alpha-toxin with the membrane, rather than specific receptor-binding. The pH during toxin binding did not influence the subsequently induced membrane damage. Non-membrane damaging alpha-toxin fragment preparations also bound firmly to the cell membranes. Upon contact with Y1 cells the fragments formed complexes of the same apparent molecular size as those generated from intact alpha-toxin. Two interpretations are possible: either the fragment oligomers are somehow defective i.e. not able to form transmembrane structures or the functional relevance of toxin oligomerization for alpha-toxin-induced membrane damage must be questioned.
Blomqvist
L
L
Department of Bacteriology, Karolinska Institutet, Stockholm, Sweden.
Thelestam
M
M
D000302:Adrenal Cortex / Q000378:metabolism; D001426:Bacterial Proteins / Q000648:ultrastructure*; D001427:Bacterial Toxins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D006460:Hemolysin Proteins; D046911:Macromolecular Substances; D008970:Molecular Weight; D011485:Protein Binding; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D046911:Macromolecular Substances; C021091:staphylococcal alpha-toxin