Characterization of two putative fibronectin-binding proteins of Clostridium perfringens.
Clostridium perfringens is a Gram-positive anaerobic pathogen that causes gas gangrene and food poisoning in humans and animals. Genomic analysis of C. perfringens strain 13 revealed that this bacterium contains two genes (CPE0737 and CPE1847) that encode putative fibronectin (Fn)-binding proteins (Fbps). These genes, named fbpA and fbpB, were found to be constitutively expressed in all three strains (13, NCTC8237, CPN50) of C. perfringens, isolated from gas gangrene of human, that were tested. Both fbpA and fbpB were cloned and His-tagged, recombinant FbpA (rFbpA) and recombinant FbpB (rFbpB) were purified by Ni(2+)-Sepharose column chromatography from transformed Escherichia coli. These recombinant Fbps were shown to bind to Fn, purified from human serum, in a ligand blotting assay. Additionally, Fn bound to these rFbps in a dose-dependent manner in an enzyme-linked immunosorbent assay. Furthermore, it was found that pre-incubation of Fn with either rFbpA or rFbpB inhibited the binding of Fn to C. perfringens cells.
Katayama
Seiichi
S
Department of Life Science, Faculty of Science, Okayama University of Science, 1-1 Ridai-cho, Okayama-shi 700-0005, Japan. katayama@dls.ous.ac.jp
Nozu
Nanami
N
Okuda
Masaya
M
Hirota
Shinsuke
S
Yamasaki
Tsutomu
T
Hitsumoto
Yasuo
Y
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003016:Clostridium perfringens / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000378:metabolism*; D005738:Gas Gangrene / Q000382:microbiology; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D005353:Fibronectins; D011994:Recombinant Proteins