Identification and characterization of a fibronectin-binding protein from Clostridium difficile.
A 68 kDa fibronectin-binding protein (Fbp68) from Clostridium difficile displaying significant homology to several established or putative Fbps from other bacteria was identified. The one-copy gene is highly conserved in C. difficile isolates. Fbp68 was expressed in Escherichia coli in fusion with glutathione S-transferase; the fusion protein and the native Fbp68 were purified. Immunoblot analysis and cell fractionation experiments revealed that Fbp68 is present on the surface of the bacteria. Far-immuno dot-blotting demonstrated that Fbp68 was capable of fixing fibronectin. Indirect immunofluorescence and ELISA were employed to demonstrate that C. difficile could bind both soluble and immobilized fibronectin. With competitive adherence inhibition assays it was shown that antibodies raised against Fbp68 partially inhibited attachment of C. difficile to fibronectin and Vero cells. Furthermore, Vero cells could fix purified membrane-immobilized Fbp68. Thus Fbp68 appears to be one of the several adhesins identified to date in C. difficile.
Hennequin
Claire
C
Département de Microbiologie, Faculté de Pharmacie, Université Paris-Sud, 5 rue JB Clément, 92296 Châtenay-Malabry cedex, France.
Janoir
Claire
C
Département de Microbiologie, Faculté de Pharmacie, Université Paris-Sud, 5 rue JB Clément, 92296 Châtenay-Malabry cedex, France.
Barc
Marie-Claude
MC
Département de Microbiologie, Faculté de Pharmacie, Université Paris-Sud, 5 rue JB Clément, 92296 Châtenay-Malabry cedex, France.
Collignon
Anne
A
Département de Microbiologie, Faculté de Pharmacie, Université Paris-Sud, 5 rue JB Clément, 92296 Châtenay-Malabry cedex, France.
Karjalainen
Tuomo
T
Département de Microbiologie, Faculté de Pharmacie, Université Paris-Sud, 5 rue JB Clément, 92296 Châtenay-Malabry cedex, France.
D018829:Adhesins, Bacterial / Q000032:analysis*; D000595:Amino Acid Sequence; D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D002352:Carrier Proteins / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D002522:Chlorocebus aethiops; D016360:Clostridioides difficile / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D005353:Fibronectins / Q000502:physiology; D008969:Molecular Sequence Data; D014709:Vero Cells
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D005353:Fibronectins; C078635:fibronectin-binding proteins, bacterial