The pore-forming toxin proaerolysin is activated by furin.
Aerolysin is secreted as an inactive dimeric precursor by the bacterium Aeromonas hydrophila. Proteolytic cleavage within a mobile loop near the C terminus of the protoxin is required for oligomerization and channel formation. This loop contains the sequence KVRRAR432, which should be recognized by mammalian proprotein convertases such as furin, PACE4, and PC5/6A. Here we show that these three proteases cleave proaerolysin after Arg-432 in vitro, yielding active toxin. We also investigated the potential role of these enzymes in the in vivo activation of the protoxin. We found that Chinese hamster ovary cells were able to convert the protoxin to aerolysin in the absence of exogenous proteases and that activation did not require internalization of the toxin. The furin inhibitor alpha1-antitrypsin Portland reduced the rate of proaerolysin activation in vivo, and proaerolysin processing was even further reduced in furin-deficient FD11 Chinese hamster ovary cells. The cells were also less sensitive to proaerolysin than wild type cells; however, transient transfection of FD11 cells with the cDNA encoding furin conferred normal sensitivity to the protoxin. Together these findings argue that furin catalyzes the cell-surface activation of proaerolysin in vivo.
Abrami
L
L
Department of Biochemistry, University of Geneva, 30 Quai E. Ansermet, 1211 Geneva, Switzerland.
Fivaz
M
M
Decroly
E
E
Seidah
N G
NG
Jean
F
F
Thomas
G
G
Leppla
S H
SH
Buckley
J T
JT
van der Goot
F G
FG
D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D045683:Furin; D006460:Hemolysin Proteins / Q000378:metabolism*; D006358:Hot Temperature; D052899:Pore Forming Cytotoxic Proteins; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000494:pharmacology; D013381:Subtilisins / Q000494:pharmacology*; D014617:Vacuoles; D000515:alpha 1-Antitrypsin / Q000494:pharmacology
D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D052899:Pore Forming Cytotoxic Proteins; D011994:Recombinant Proteins; D000515:alpha 1-Antitrypsin; C107096:proaerolysin; D013381:Subtilisins; D045683:Furin