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title Aeromonas piscicola AH-3 expresses an extracellular collagenase with cytotoxic properties
authors Duarte, AS; Cavaleiro, E; Pereira, C; Merino, S; Esteves, AC; Duarte, EP; Tomas, JM; Correia, AC
author full name Duarte, A. S.; Cavaleiro, E.; Pereira, C.; Merino, S.; Esteves, A. C.; Duarte, E. P.; Tomas, J. M.; Correia, A. C.
title Aeromonas piscicola AH-3 expresses an extracellular collagenase with cytotoxic properties
nationality internacional
source LETTERS IN APPLIED MICROBIOLOGY
language English
document type Article
author keywords AH-3; collagen interaction; cytotoxicity; metalloprotease; microbial collagenase
keywords plus BIOCHEMICAL-CHARACTERIZATION; CLOSTRIDIAL COLLAGENASES; DUPUYTREN DISEASE; ESCHERICHIA-COLI; GENOME SEQUENCE; GENE-EXPRESSION; VIRULENCE; INJECTION; ENZYMES; BINDING
abstract The aim of this study was to investigate the presence and the phenotypic expression of a gene coding for a putative collagenase. This gene (AHA_0517) was identified in Aeromonas hydrophila ATCC 7966 genome and named colAh. We constructed and characterized an Aeromonas piscicola AH-3::colAh knockout mutant. Collagenolytic activity of the wild-type and mutant strains was determined, demonstrating that colAh encodes for a collagenase. ColAh-collagen interaction was assayed by Far-Western blot, and cytopathic effects were investigated in Vero cells. We demonstrated that ColAh is a gluzincin metallopeptidase (approx. 100 kDa), able to cleave and physically interact with collagen, that contributes for Aeromonas collagenolytic activity and cytotoxicity. ColAh possess the consensus HEXXH sequence and a glutamic acid as the third zinc binding positioned downstream the HEXXH motif, but has low sequence similarity and distinct domain architecture to the well-known clostridial collagenases. In addition, these results highlight the importance of exploring new microbial collagenases that may have significant relevance for the health and biotechnological industries.
author address [Duarte, A. S.; Cavaleiro, E.; Pereira, C.; Esteves, A. C.; Correia, A. C.] Univ Aveiro, Dept Biol, P-3810193 Aveiro, Portugal; [Duarte, A. S.; Cavaleiro, E.; Pereira, C.; Esteves, A. C.; Correia, A. C.] Univ Aveiro, CESAM, P-3810193 Aveiro, Portugal; [Pereira, C.; Merino, S.; Tomas, J. M.] Univ Barcelona, Fac Biol, Dept Microbiol, Barcelona, Spain; [Duarte, E. P.] Univ Coimbra, Ctr Neurosci & Cell Biol, Coimbra, Portugal; [Duarte, E. P.] Univ Coimbra, Dept Zool, Coimbra, Portugal
reprint address Duarte, AS (reprint author), Univ Aveiro, Dept Biol, P-3810193 Aveiro, Portugal.
e-mail address asduarte@ua.pt
researcherid number Correia, Antonio/B-1593-2008; Cavaleiro, Eliana Marisa S./A-2957-2012; Esteves, Ana Cristina/B-2939-2008
orcid number Correia, Antonio/0000-0002-5115-1429; Cavaleiro, Eliana Marisa S./0000-0002-3862-915X; Esteves, Ana Cristina/0000-0003-2239-2976
funding agency and grant number European Funds through COMPETE; National Funds through the Portuguese Science Foundation (FCT) [PEst-C/MAR/LA0017/2013]; FCT [BPD/38008/2007, BPD/46290/2008, BD/47502/2008]; Plan Nacional de I+D+I grant (Ministerio de Educacion, Ciencia y Deporte and Ministerio de Sanidad, Spain); FIS grant (Ministerio de Educacion, Ciencia y Deporte and Ministerio de Sanidad, Spain); Generalitat de Catalunya (Centre de Referencia en Biotecnologia)
funding text This work was supported by European Funds through COMPETE and by National Funds through the Portuguese Science Foundation (FCT) within project PEst-C/MAR/LA0017/2013. The author also wish to acknowledge FCT for grants to AC Esteves, AS Duarte and E Cavaleiro (FCT; BPD/38008/2007, BPD/46290/2008 and BD/47502/2008). Part of this work was also supported by Plan Nacional de I+D+I and FIS grants (Ministerio de Educacion, Ciencia y Deporte and Ministerio de Sanidad, Spain) and by Generalitat de Catalunya (Centre de Referencia en Biotecnologia).
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cited reference count 62
publisher WILEY-BLACKWELL
publisher city HOBOKEN
publisher address 111 RIVER ST, HOBOKEN 07030-5774, NJ USA
issn 0266-8254
29-character source abbreviation LETT APPL MICROBIOL
iso source abbreviation Lett. Appl. Microbiol.
publication date MAR
year published 2015
volume 60
issue 3
beginning page 288
ending page 297
digital object identifier (doi) 10.1111/lam.12373
page count 10
web of science category Biotechnology & Applied Microbiology; Microbiology
subject category Biotechnology & Applied Microbiology; Microbiology
document delivery number CB4UF
unique article identifier WOS:000349623000013
link http://onlinelibrary.wiley.com/doi/10.1111/lam.12373/abstract;jsessionid=42E7EACE8016E32AD9BC2C73B62E0717.f04t01