title: Procaspase-3 activation by a metalloprotease secreted from Vibrio vulnificus.
abstract: Vibrio vulnificus is a marine bacterium and a human pathogen capable of causing wound infection and septicemia. We previously showed that the metalloprotease vEP secreted by V. vulnificus activates prothrombin in vitro. To further investigate the ability of vEP to activate other zymogens, we used a mutant form of procaspase-3 which lacks the native cleavage sites as a zymogen. The mutant zymogen was activated by vEP to yield a mature enzyme with a maximum increase in caspase-3 activity of approximately 14-fold in a time-dependent manner. However, the increase started to decline with prolonged incubation and with higher protease concentration as a result of further cleavage of the mature enzyme. Western blot analysis revealed a band of approximately 17 kDa for the cleavage product, which corresponded with the change in caspase-3 activity. The activated procaspase-3 by vEP was also able to cleave poly(ADP-ribose) polymerase in a cell-free system, and was inhibited by Ac-DEVD-CHO, a potent caspase-3 inhibitor. The results presented are the first to demonstrate the in vitro activation of one of the crucial enzymes involved in cell death by a bacterial extracellular metalloprotease.
authors.affiliation: Research Center for Proteineous Materials, Chosun University, Gwangju 501-759, Korea.
authors.forename: Hyo Young
authors.initials: HY
authors.lastname: Kim
authors.forename: Alan K
authors.initials: AK
authors.lastname: Chang
authors.forename: Jung Eun
authors.initials: JE
authors.lastname: Park
authors.forename: Il-Seon
authors.initials: IS
authors.lastname: Park
authors.forename: Seong Myeong
authors.initials: SM
authors.lastname: Yoon
authors.forename: Jung Sup
authors.initials: JS
authors.lastname: Lee
mesh_terms: D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D045726:Metalloproteases / Q000378:metabolism* / Q000494:pharmacology; D010446:Peptide Fragments / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D013997:Time Factors; D041261:Vibrio vulnificus / Q000201:enzymology* / Q000378:metabolism