Biological relevance of natural alpha-toxin fragments from Staphylococcus aureus.
Serine proteases represent an essential part of cellular homeostasis by generating biologically active peptides. In bacteria, proteolysis serves two different roles: a major housekeeping function and the destruction of foreign or target cell proteins, thereby promoting bacterial invasion. In the process, other virulence factors such as exotoxins become affected. In Staphylococcus aureus culture supernatant, the pore-forming alpha-toxin is cleaved by the coexpressed V8 protease and aureolysin. The oligomerizing and pore-forming abilities of five such spontaneously occurring N- and C-terminal alpha-toxin fragments were studied. (3)H-marked alpha-toxin fragments bound to rabbit erythrocyte membranes but only fragments with intact C termini, missing 8, 12 and 71 amino acids from their N-terminal, formed stable oligomers. All isolated fragments induced intoxication of mouse adrenocortical Y1 cells in vitro, though the nature of membrane damage for a fragment, degraded at its C terminus, remained obscure. Only one fragment, missing the first eight N-terminal amino acids, induced irreversible intoxication of Y1 cells in the same manner as the intact toxin. Four of the isolated fragments caused swelling, indicating altered channel formation. Fragments missing 12 and 71 amino acids from the N terminus occupied the same binding sites on Y1 cell membranes, though they inhibited membrane damage caused by intact toxin. In conclusion, N-terminal deletions up to 71 amino acids are tolerated, though the kinetics of channel formation and the channel's properties are altered. In contrast, digestion at the C terminus results in nonfunctional species.
Kwak
Young-Keun
YK
Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden. Young-Keun.Kwak@ki.se
Högbom
Martin
M
Colque-Navarro
Patricia
P
Möllby
Roland
R
Vécsey-Semjén
Beatrix
B
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D001427:Bacterial Toxins / Q000378:metabolism*; D002460:Cell Line; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D008666:Metalloendopeptidases / Q000378:metabolism; D051379:Mice; D011817:Rabbits; D013211:Staphylococcus aureus / Q000254:growth & development / Q000378:metabolism*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D008666:Metalloendopeptidases; C115504:aureolysin