Protein F: an adhesin of Streptococcus pyogenes binds fibronectin via two distinct domains.
The binding of Streptococcus pyogenes to fibronectin (FN) enables the adherence of this pathogen to target epithelial cells, which is the first necessary step for initiation of infection. Binding is mediated by a bacterial surface protein termed protein F. Here we provide the complete structure of protein F and identify two domains responsible for binding to fibronectin. The first domain is located towards the C-terminal end of the molecule and is composed of five repeats of 37 amino acids that are completely repeated four times and a fifth time partially. The second domain is adjacent to the first domain and is located on the N-terminal side of it. It is composed of a single stretch of 43 amino acids. Protein F expressed in Escherichia coli completely blocked the binding of fibronectin to S. pyogenes. However, mutant proteins that contained only one or the other of the two domains were only capable of partial blockage of binding. Complete blockage of binding of fibronectin could be achieved when a protein extract containing the N-terminal domain was mixed in a binding reaction with a protein extract containing the C-terminal domain. Similarly, a purified recombinant protein containing the two domains only, blocked the binding completely. In contrast, a purified recombinant protein containing just the C-terminal domain, blocked the binding partially. A clone exclusively expressing the C-terminal domain, completely blocked the binding of the 30 kDa N-terminal fragment of fibronectin to S. pyogenes, whereas a clone expressing the N-terminal domain failed to block the binding of this FN fragment.(ABSTRACT TRUNCATED AT 250 WORDS)
Sela
S
S
Department of Clinical Microbiology, Hebrew University Hadassah Medical School, Jerusalem, Israel.
Aviv
A
A
Tovi
A
A
Burstein
I
I
Caparon
M G
MG
Hanski
E
E
D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005353:Fibronectins / Q000378:metabolism*; D005798:Genes, Bacterial; D006094:Gram-Positive Bacteria / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
D018829:Adhesins, Bacterial; D017931:DNA Primers; D004269:DNA, Bacterial; D005353:Fibronectins; C078635:fibronectin-binding proteins, bacterial