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Microbiology 144 (1998), 119-126; DOI  10.1099/00221287-144-1-119
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Protein F, a fibronectin-binding protein of Streptococcus pyogenes, also binds human fibrinogen: isolation of the protein and mapping of the binding region

Viacheslav Katerov1,2, Andrej Andreev1, Claes Schalén2,1 and Artem A. Totolian1

Institute of Experimental Medicine, Academy of the Medical Sciences, St Petersburg, Russia
Department of Medical Microbiology, University of Lund, Sölvegatan 23, S-22362 Lund, Sweden

ABSTRACT

Summary: During screening of a gene library of Streptococcus pyogenes type M15 for fibrinogen-binding material, a protein of approximately 100 kDa, encoded outside the vir region, was found. DNA sequencing revealed this component to be identical to protein F, a fibronectin-binding protein. Isolation of the recombinant protein, termed F15, was performed by the use of fibrinogen affinity chromatography. The affinity constant (Ka) of protein F15 for fibrinogen, 1.25 x 107 mol-1, was lower than that for fibronectin, 1.8 x 108 mol-1. The fibrinogen-binding domain was located in the N-terminal part of the molecule, while the fibronectin-binding domains, as previously determined, were in the C-terminal portion of protein F. To examine the amino acid sequence heterogeneity of protein F, the 5' part of the prtF gene, corresponding to the N-terminal variable region of the protein, was amplified by PCR from 12 strains of S. pyogenes belonging to six different M-types. Alignment of these nucleotide sequences indicated that the 5' portion of the prtF gene had probably undergone a number of intragenic recombination and horizontal gene transfer events, allowing a pattern of structural diversity of protein F observed earlier for some other streptococcal virulence factors. There was no strict correlation between M-type and nucleotide sequence of the variable region of the prtF gene and, compared to streptococcal M protein, the overall variation observed for protein F appeared more limited.

Author for correspondence: Claes Schalén. Tel: + 46 46173284. Fax: +46 46189117.

Keywords: Streptococcus pyogenes, fibrinogen, fibronectin, protein F




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