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Infection and Immunity, July 2006, p. 4083-4093, Vol. 74, No. 7
0019-9567/06/$08.00+0 doi:10.1128/IAI.00297-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.
Burkholderia cenocepacia ZmpB Is a Broad-Specificity Zinc Metalloprotease Involved in Virulence
C. Kooi,
B. Subsin,
R. Chen,
B. Pohorelic, and
P. A. Sokol*
Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada T2N 4N1
Received 22 February 2006/
Returned for modification 5 April 2006/
Accepted 18 April 2006
In previous studies we characterized the Burkholderia cenocepacia ZmpA zinc metalloprotease. In this study, we determined that B. cenocepacia has an additional metalloprotease, which we designated ZmpB. The zmpB gene is present in the same species as zmpA and was detected in B. cepacia, B. cenocepacia, B. stabilis, B. ambifaria, and B. pyrrocinia but was absent from B. multivorans, B. vietnamiensis, B. dolosa, and B. anthina. The zmpB gene was expressed, and ZmpB was purified from Escherichia coli by using the pPROEXHTa His6 Tag expression system. ZmpB has a predicted preproenzyme structure typical of thermolysin-like proteases and is distantly related to Bacillus cereus bacillolysin. ZmpB was expressed as a 63-kDa preproenzyme precursor that was autocatalytically cleaved into mature ZmpB (35 kDa) and a 27-kDa prepropeptide. EDTA, 1,10-phenanthroline, and Zn2+ cations inhibited ZmpB enzyme activity, indicating that it is a metalloprotease. ZmpB had proteolytic activity against
-1 proteinase inhibitor,
2-macrogobulin, type IV collagen, fibronectin, lactoferrin, transferrin, and immunoglobulins. B. cenocepacia zmpB and zmpA zmpB mutants had no proteolytic activity against casein and were less virulent in a rat agar bead chronic infection model, indicating that zmpB is involved in B. cenocepacia virulence. Expression of zmpB was regulated by both the CepIR and CciIR quorum-sensing systems.
* Corresponding author. Mailing address: Department of Microbiology and Infectious Diseases, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada T2N 4N1. Phone: (403) 220-6037. Fax: (403) 270-2772. E-mail:
psokol{at}ucalgary.ca.
Editor: J. T. Barbieri
These authors contributed equally to this study.
Infection and Immunity, July 2006, p. 4083-4093, Vol. 74, No. 7
0019-9567/06/$08.00+0 doi:10.1128/IAI.00297-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.
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