Cloning, sequencing, and expression of a fibronectin/fibrinogen-binding protein from group A streptococci

Harry Courtney, Y. Li, James Dale, D. L. Hasty

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Abstract

Lipoteichoic acid and several streptococcal proteins have been reported to bind fibronectin (Fn) or fibrinogen (Fgn), which may serve as host receptors. We searched for such proteins by screening a library of genes from M type 5 group A streptococci cloned into Escherichia coli. Lysates of clones were probed with biotinylated Fn and biotinylated Fgn. One clone expressed a 54- kDa protein that reacted with Fn and Fgn. The protein, termed FBP54, was purified and used to immunize rabbits. Anti-FBP54 serum reacted with purified, recombinant FBP54 and with a protein of similar electrophoretic mobility in extracts of M type 5, 6, and 24 streptococci. Anti-FBP54 serum also reacted with 5 of 15 strains of intact, live streptococci, suggesting that FBP54 may be a surface antigen. Southern blot analysis confirmed that the gene is found in group A streptococci but not in Staphylococcus aureus or E. coli. The cloned gene was sequenced and contained an open reading frame encoding a protein with a calculated molecular weight of 54,186. Partial amino acid sequencing of purified FBP54 confirmed that this open reading frame encoded the protein. As determined by utilizing fusion proteins containing truncated forms of FBP54, the primary Fn/Fgn-binding domain appears to be contained in residues 1 to 89. These data suggest that FBP54 may be a surface protein of streptococci that reacts with both Fn and Fgn and therefore may participate in the adhesion of group A streptococci to host cells.

Original languageEnglish (US)
Pages (from-to)3937-3946
Number of pages10
JournalInfection and Immunity
Volume62
Issue number9
StatePublished - Jan 1 1994

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Streptococcus
Fibronectins
Fibrinogen
Organism Cloning
Carrier Proteins
Proteins
Open Reading Frames
Clone Cells
Escherichia coli
Protein Sequence Analysis
Surface Antigens
Southern Blotting
Serum
Gene Library
Genes
Staphylococcus aureus
Membrane Proteins
Molecular Weight
Rabbits

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

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Cloning, sequencing, and expression of a fibronectin/fibrinogen-binding protein from group A streptococci. / Courtney, Harry; Li, Y.; Dale, James; Hasty, D. L.

In: Infection and Immunity, Vol. 62, No. 9, 01.01.1994, p. 3937-3946.

Research output: Contribution to journalArticle

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