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By screening a genomic lambda library of Streptococcus equi subsp. zooepidemicus, we have cloned and sequenced a gene, termed fnz, encoding a fibronectin (Fn)-binding protein called FNZ. On the basis of the deduced amino acid sequence of FNZ, the mature protein has a molecular mass of approximately 61 kDa. Analysis of FNZ reveals a structural organization similar to that of other cell surface proteins from streptococci and staphylococci. The Fn-binding activity is localized to two domains in the C-terminal part of FNZ. One domain is composed of five repeats, which contain a motif similar to what has earlier been found in other Fn-binding proteins in streptococci and staphylococci. The first and second repeats are separated by a short stretch of amino acids, including the motif LAGESGET, which is an important part of the second Fn-binding domain. This motif is also present in an Fn-binding domain (UR) in protein F of Streptococcus pyogenes. A fusion protein covering the Fn-binding domain of FNZ inhibits the binding of the 29-kDa N-terminal fragment of Fn to cells of various streptococcal species as well as to Staphylococcus aureus.

Genetic diversity of the streptokinase gene (sk) from 36 strains of S. equisimilis and 54 strains of group G streptococci was examined. The strains were isolated from patients with various streptococcal disease manifestations and healthy carriers. The region of the gene that corresponds to amino acid residues 174-244, was PCR amplified. The amplified product was subjected to MluI, PvuII, DraI and DdeI digestion. Based on the restriction enzyme digestion patterns nine sk alleles were recognized. There was no correlation between the various sk gene alleles and streptococcal disease manifestations. Three of the nine sk gene alleles, sk4, sk7, and sk8, were detected earlier among group A streptococci. The other six alleles were unique to S. equisimilis and group G streptococci. The most common alleles were sk5, found in 21/90 (23%) and sk10 detected in 43/90 (47%) of the strains. Alleles sk1 and sk2, the most frequent among group A streptococci, were not found among the strains in the present investigation. Thus, it appears that the sk gene has been evolving in line with other species distinguishing features of the streptococci.

This work describes the cloning and sequencing of a gene encoding a plasma protein receptor from Streptococcus zooepidemicus. This receptor, termed protein ZAG, is a 45-kDa protein that binds alpha 2-macroglobulin (alpha 2M), serum albumin, and immunoglobulin G (IgG). The IgG-binding activity is located in the C-terminal part of the molecule and is mediated by two repeated domains highly homologous to each other as well as to the corresponding domains in streptococcal type III Fc receptors. The IgG-binding profile of protein ZAG is similar to that previously reported for S. zooepidemicus. Binding to serum albumin is mediated by a short amino acid sequence in the middle of the molecule. This domain shows homology to previously described albumin-binding proteins from streptococci, and the albumin-binding profile of protein ZAG is similar to that of streptococcal protein G. The N-terminal part of protein ZAG, which mediates binding to the plasma proteinase inhibitor alpha 2M, is composed of a unique stretch of amino acids. Protein ZAG competes for the same, or nearby, binding site(s) in alpha 2M as do two recently described Streptococcus dysgalactiae receptors, although the sequences of the alpha 2M-binding domains in these three receptors show only minor sequence similarities.

The genetic structure of beta-haemolytic Lancefield group C streptococci isolated from horses in Australia was examined by multilocus enzyme electrophoresis. The 249 isolates comprised 70 classified phenotypically as Streptococcus equi subspecies equi, 177 classified as S equi subspecies zooepidemicus and two which were unclassifiable. Forty-one electrophoretic types were identified which could be classified into three major clusters, A, B and C. Of the isolates, 178 fell into cluster B (types 4 to 22) and lay within a genetic distance of 0.36. Sixty-nine of the 70 S equi subspecies equi isolates fell into type 12, which suggests that they were members of a single clone, and the isolates from abscesses were significantly more likely to belong to type 12 than those from horses with no clinical signs (P < 0.001). There were no other significant associations between electrophoretic types or clusters and the isolation of the organism from particular sites. These data suggested that S zooepidemicus may be the archetypal species from which the clone designated subspecies equi has been derived. If isolates of the subspecies equi from other geographical regions also prove to be members of electrophoretic type 12, this hypothesis would be strengthened.