Characterization of the collagen-like exosporium protein, BclA1, of Clostridium difficile spores

Marjorie Pizarro-Guajardo, Valeria Olguín-Araneda, Jonathan Barra-Carrasco, Christian Brito-Silva, Mahfuzur R. Sarker, Daniel Paredes-Sabja

Resultado de la investigación: Article

29 Citas (Scopus)

Resumen

Spores of Clostridium difficile are essential for infection, persistence and transmission of C.difficile infections (CDI). Proteins of the surface of C.difficile spores are thought to be essential for initiation and persistence of CDI. In this work, we demonstrate that three C. difficile collagen-like exosporium proteins (BclA) encoded in the C.difficile 630 genome are expressed during sporulation and localize to the spore via their N-terminal domains. Using polyclonal antibodies against the N- and C-terminal domains and full length BclA1 we demonstrate that BclA1 is likely to be localized to the exosporium layer, presumably undergoes post-translational cleavages and might be cross-linked with other exosporium proteins. The collagen-like region of recombinant BclA1 and BclA2 was susceptible to collagenase degradation. Collagenase digestion assay of C.difficile spores suggests that, similarly as in Bacillus anthracis BclA, the N-terminal domain and the C-terminal domain of BclA1 might be buried in the basal layer and oriented to the exosporium surface, respectively. We also demonstrate that the collagen-like BclAs proteins do not contribute to the spore hydrophobicity and its absence slightly increased the adherence of spores to Caco-2 cells. BclA1 was also shown to have poor immunogenic properties. These results provide the first study on the BclA1 collagen-like proteins of C.difficile spores.

Idioma originalEnglish
Páginas (desde-hasta)18-30
Número de páginas13
PublicaciónAnaerobe
Volumen25
DOI
EstadoPublished - 1 ene 2014

Huella dactilar

Clostridium difficile
Spores
Collagen
Proteins
Collagenases
Protein C
Bacillus anthracis
Infectious Disease Transmission
Caco-2 Cells
Infection
Hydrophobic and Hydrophilic Interactions
Digestion
Genome
Antibodies

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Citar esto

Pizarro-Guajardo, M., Olguín-Araneda, V., Barra-Carrasco, J., Brito-Silva, C., Sarker, M. R., & Paredes-Sabja, D. (2014). Characterization of the collagen-like exosporium protein, BclA1, of Clostridium difficile spores. Anaerobe, 25, 18-30. https://doi.org/10.1016/j.anaerobe.2013.11.003
Pizarro-Guajardo, Marjorie ; Olguín-Araneda, Valeria ; Barra-Carrasco, Jonathan ; Brito-Silva, Christian ; Sarker, Mahfuzur R. ; Paredes-Sabja, Daniel. / Characterization of the collagen-like exosporium protein, BclA1, of Clostridium difficile spores. En: Anaerobe. 2014 ; Vol. 25. pp. 18-30.
@article{aa6fe0a3c402448b8dcfa4c5fa5dfea2,
title = "Characterization of the collagen-like exosporium protein, BclA1, of Clostridium difficile spores",
abstract = "Spores of Clostridium difficile are essential for infection, persistence and transmission of C.difficile infections (CDI). Proteins of the surface of C.difficile spores are thought to be essential for initiation and persistence of CDI. In this work, we demonstrate that three C. difficile collagen-like exosporium proteins (BclA) encoded in the C.difficile 630 genome are expressed during sporulation and localize to the spore via their N-terminal domains. Using polyclonal antibodies against the N- and C-terminal domains and full length BclA1 we demonstrate that BclA1 is likely to be localized to the exosporium layer, presumably undergoes post-translational cleavages and might be cross-linked with other exosporium proteins. The collagen-like region of recombinant BclA1 and BclA2 was susceptible to collagenase degradation. Collagenase digestion assay of C.difficile spores suggests that, similarly as in Bacillus anthracis BclA, the N-terminal domain and the C-terminal domain of BclA1 might be buried in the basal layer and oriented to the exosporium surface, respectively. We also demonstrate that the collagen-like BclAs proteins do not contribute to the spore hydrophobicity and its absence slightly increased the adherence of spores to Caco-2 cells. BclA1 was also shown to have poor immunogenic properties. These results provide the first study on the BclA1 collagen-like proteins of C.difficile spores.",
keywords = "BclA, Clostridium difficile, Exosporium, Spore",
author = "Marjorie Pizarro-Guajardo and Valeria Olgu{\'i}n-Araneda and Jonathan Barra-Carrasco and Christian Brito-Silva and Sarker, {Mahfuzur R.} and Daniel Paredes-Sabja",
year = "2014",
month = "1",
day = "1",
doi = "10.1016/j.anaerobe.2013.11.003",
language = "English",
volume = "25",
pages = "18--30",
journal = "Anaerobe",
issn = "1075-9964",
publisher = "Academic Press Inc.",

}

Pizarro-Guajardo, M, Olguín-Araneda, V, Barra-Carrasco, J, Brito-Silva, C, Sarker, MR & Paredes-Sabja, D 2014, 'Characterization of the collagen-like exosporium protein, BclA1, of Clostridium difficile spores', Anaerobe, vol. 25, pp. 18-30. https://doi.org/10.1016/j.anaerobe.2013.11.003

Characterization of the collagen-like exosporium protein, BclA1, of Clostridium difficile spores. / Pizarro-Guajardo, Marjorie; Olguín-Araneda, Valeria; Barra-Carrasco, Jonathan; Brito-Silva, Christian; Sarker, Mahfuzur R.; Paredes-Sabja, Daniel.

En: Anaerobe, Vol. 25, 01.01.2014, p. 18-30.

Resultado de la investigación: Article

TY - JOUR

T1 - Characterization of the collagen-like exosporium protein, BclA1, of Clostridium difficile spores

AU - Pizarro-Guajardo, Marjorie

AU - Olguín-Araneda, Valeria

AU - Barra-Carrasco, Jonathan

AU - Brito-Silva, Christian

AU - Sarker, Mahfuzur R.

AU - Paredes-Sabja, Daniel

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Spores of Clostridium difficile are essential for infection, persistence and transmission of C.difficile infections (CDI). Proteins of the surface of C.difficile spores are thought to be essential for initiation and persistence of CDI. In this work, we demonstrate that three C. difficile collagen-like exosporium proteins (BclA) encoded in the C.difficile 630 genome are expressed during sporulation and localize to the spore via their N-terminal domains. Using polyclonal antibodies against the N- and C-terminal domains and full length BclA1 we demonstrate that BclA1 is likely to be localized to the exosporium layer, presumably undergoes post-translational cleavages and might be cross-linked with other exosporium proteins. The collagen-like region of recombinant BclA1 and BclA2 was susceptible to collagenase degradation. Collagenase digestion assay of C.difficile spores suggests that, similarly as in Bacillus anthracis BclA, the N-terminal domain and the C-terminal domain of BclA1 might be buried in the basal layer and oriented to the exosporium surface, respectively. We also demonstrate that the collagen-like BclAs proteins do not contribute to the spore hydrophobicity and its absence slightly increased the adherence of spores to Caco-2 cells. BclA1 was also shown to have poor immunogenic properties. These results provide the first study on the BclA1 collagen-like proteins of C.difficile spores.

AB - Spores of Clostridium difficile are essential for infection, persistence and transmission of C.difficile infections (CDI). Proteins of the surface of C.difficile spores are thought to be essential for initiation and persistence of CDI. In this work, we demonstrate that three C. difficile collagen-like exosporium proteins (BclA) encoded in the C.difficile 630 genome are expressed during sporulation and localize to the spore via their N-terminal domains. Using polyclonal antibodies against the N- and C-terminal domains and full length BclA1 we demonstrate that BclA1 is likely to be localized to the exosporium layer, presumably undergoes post-translational cleavages and might be cross-linked with other exosporium proteins. The collagen-like region of recombinant BclA1 and BclA2 was susceptible to collagenase degradation. Collagenase digestion assay of C.difficile spores suggests that, similarly as in Bacillus anthracis BclA, the N-terminal domain and the C-terminal domain of BclA1 might be buried in the basal layer and oriented to the exosporium surface, respectively. We also demonstrate that the collagen-like BclAs proteins do not contribute to the spore hydrophobicity and its absence slightly increased the adherence of spores to Caco-2 cells. BclA1 was also shown to have poor immunogenic properties. These results provide the first study on the BclA1 collagen-like proteins of C.difficile spores.

KW - BclA

KW - Clostridium difficile

KW - Exosporium

KW - Spore

UR - http://www.scopus.com/inward/record.url?scp=84889598919&partnerID=8YFLogxK

U2 - 10.1016/j.anaerobe.2013.11.003

DO - 10.1016/j.anaerobe.2013.11.003

M3 - Article

C2 - 24269655

AN - SCOPUS:84889598919

VL - 25

SP - 18

EP - 30

JO - Anaerobe

JF - Anaerobe

SN - 1075-9964

ER -