The bactericidal action on Escherichia coli of ZF-RNase-3 is triggered by the suicidal action of the bacterium OmpT protease.
ZF-RNase-3 is one of the RNases from zebrafish (Danio rerio) with special (i.e. noncatalytic) properties. These include angiogenic and bactericidal activities. Given the interest of fish RNases as host-defense effectors, we studied the mechanism of the bactericidal action of ZF-RNase-3 on Escherichia coli as a model Gram-negative bacterium. The results obtained indicate that the bactericidal activity of ZF-RNase-3 is not lost when its catalytic RNase activity is obliterated. On the other hand, fully denatured ZF-RNase-3 conserves its bactericidal activity. When ZF-RNase-3 is added to E. coli cultures, it is cleaved at a specific Arg-Arg peptide bond, thus engendering two peptide fragments. The larger fragment (residues 31-124), produced by proteolysis and reduction of a disulfide, is recognized as the actual bactericidal agent. The protease responsible for the proteolytic attack has been identified with OmpT, an outer membrane E. coli omptin protease. However, the most remarkable result obtained in the present study is the finding that the microbicidal action of ZF-RNase-3 can be achieved only with the suicidal cooperation of the bacterium itself.
Zanfardino
Anna
A
Dipartimento di Biologia Strutturale e Funzionale, Università di Napoli Federico II, Napoli, Italy.
Pizzo
Elio
E
Di Maro
Antimo
A
Varcamonti
Mario
M
D'Alessio
Giuseppe
G
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D047150:Eosinophil Cationic Protein / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism*; D015027:Zebrafish / Q000235:genetics
D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D011994:Recombinant Proteins; D047150:Eosinophil Cationic Protein; D012697:Serine Endopeptidases; C057840:omptin outer membrane protease