Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage

Enterococci account for nearly 10% of all nosocomial infections and constitute a significant treatment challenge due to their multidrug resistance properties. One of the well-studied virulence factors of Enterococcus faecalis is a secreted bacterial protease, termed gelatinase, which has been shown...

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Autores principales: Del Papa, María Florencia, Hancock, Lynn E., Thomas, Vinai C., Perego, Marta
Formato: Articulo
Lenguaje:Inglés
Publicado: 2007
Materias:
Biología
Enterococcus faecalis
Infecciones hospitalarias
Bacteriología
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/83204
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-83204
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Biología
Enterococcus faecalis
Infecciones hospitalarias
Bacteriología
spellingShingle Biología
Enterococcus faecalis
Infecciones hospitalarias
Bacteriología
Del Papa, María Florencia
Hancock, Lynn E.
Thomas, Vinai C.
Perego, Marta
Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage
topic_facet Biología
Enterococcus faecalis
Infecciones hospitalarias
Bacteriología
description Enterococci account for nearly 10% of all nosocomial infections and constitute a significant treatment challenge due to their multidrug resistance properties. One of the well-studied virulence factors of Enterococcus faecalis is a secreted bacterial protease, termed gelatinase, which has been shown to contribute to the process of biofilm formation. Gelatinase belongs to the M4 family of bacterial zinc metalloendopeptidases, typified by thermolysin. Gelatinase is synthesized as a preproenzyme consisting of a signal sequence, a putative propeptide, and then the mature enzyme. We determined that the molecular mass of the mature protein isolated from culture supernatant was 33,030 Da, which differed from the predicted molecular mass, 34,570 Da, by over 1,500 Da. Using N-terminal sequencing, we confirmed that the mature protein begins at the previously identified sequence VGSEV, thus suggesting that the 1,500-Da molecular mass difference resulted from a C-terminal processing event. By using mutants with site-directed mutations within a predicted C-terminal processing site and mutants with C-terminal deletions fused to a hexahistidine tag, we determined that the processing site is likely to be between residues D304 and 1305 and that it requires the Q306 residue. The results suggest that the E. faecalis gelatinase requires C-terminal processing for full activation of protease activity, making it a unique enzyme among the members of the M4 family of proteases of gram-positive bacteria.
format Articulo
Articulo
author Del Papa, María Florencia
Hancock, Lynn E.
Thomas, Vinai C.
Perego, Marta
author_facet Del Papa, María Florencia
Hancock, Lynn E.
Thomas, Vinai C.
Perego, Marta
author_sort Del Papa, María Florencia
title Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage
title_short Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage
title_full Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage
title_fullStr Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage
title_full_unstemmed Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage
title_sort full activation of enterococcus faecalis gelatinase by a c-terminal proteolytic cleavage
publishDate 2007
url http://sedici.unlp.edu.ar/handle/10915/83204
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AT hancocklynne fullactivationofenterococcusfaecalisgelatinasebyacterminalproteolyticcleavage
AT thomasvinaic fullactivationofenterococcusfaecalisgelatinasebyacterminalproteolyticcleavage
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bdutipo_str Repositorios
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