Vibrio cholerae hemagglutinin/protease (HA/protease) causes morphological changes in cultured epithelial cells and perturbs their paracellular barrier function.
In this report, we describe the cytotoxic activity of the cholera hemagglutinin/protease (HA/protease). A concentrated protein sample from the 37 degrees C overnight culture supernatant of CVD110, a delta ctxA, delta zot, delta Ace and hlyA::(ctxB mer) mutant of El Tor biotype Ogawa serotype strain E7946 caused morphological changes in cultured MDCK-I epithelial cells and altered their arrangement of filamentous actin (F-actin) and Zonula occludens-associated protein ZO-1. The drastic morphological changes can be inhibited by Zincov, a specific bacterial metalloprotease inhibitor. The cytotoxic fractions of the sample after FPLC gelfiltration fractionation showed two visible protein bands with molecular weights of approximately 34- and 32 kDa. Microsequencing of these two proteins revealed that they were the cholera HA/protease.
Wu
Z
Z
Department of Medical Microbiology, Linköping University, Sweden.
Milton
D
D
Nybom
P
P
Sjö
A
A
Magnusson
K E
KE
D000199:Actins / Q000187:drug effects; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D002771:Cholera / Q000209:etiology; D004285:Dogs; D017097:Electric Impedance; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects; D006388:Hemagglutinins / Q000633:toxicity*; D007668:Kidney / Q000166:cytology; D008565:Membrane Proteins / Q000187:drug effects; D008666:Metalloendopeptidases / Q000633:toxicity*; D010539:Permeability / Q000187:drug effects; D010750:Phosphoproteins / Q000187:drug effects; D014734:Vibrio cholerae / Q000472:pathogenicity; D062826:Zonula Occludens-1 Protein
D000199:Actins; D006388:Hemagglutinins; D008565:Membrane Proteins; D010750:Phosphoproteins; D062826:Zonula Occludens-1 Protein; D008666:Metalloendopeptidases; C039814:hemagglutinin-protease