Expression of protective and cardiac tissue cross-reactive epitopes of type 5 streptococcal M protein in Escherichia coli

T. P. Poirier, M. A. Kehoe, James Dale, K. N. Timmis, E. H. Beachey

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Abstract

The immunochemical properties of type 5 M protein antigens that were expressed in Escherichia coli K-12 by recombinant lambda bacteriophages isolated from a gene bank of serotype 5 Streptococcus pyogenes have been analyzed in detail. M proteins from partially purified bacteriophage lysates displayed precipitin lines of identity with a purified peptic extract of type 5 M protein (pep M5) in immunodiffusion assays. Immunoblot analyses of the M protein-positive lysates demonstrated that the cloned M protein component resided in five polypeptides with relative molecular weights of 57,900 (57.9K), 55.4K, 52.9K, 40.0K, and 32.6K. The hybrid λ phage (λM5) produced M protein contained immunoprotective epitopes; λM5 protein inhibited opsonization of type 5 streptococci by pep M5 antibodies, and antiserum raised against λM5 lysates opsonized type 5 streptococci. Each of the five antigenic polypeptides of te recombinant phage M protein also shared epitopes with human heart tissue, as demonstrated by the reactivity of immunoblots of λM5 antigens separated on sodium dodecyl sulfate gels with anti-pep M5 antibodies absorbed to and eluted from human heart sarcolemmal membranes. Moreover, antiserum raised against the λM5 lysates reacted with sarcolemmal membrane proteins with relative molecular weights of 200K, 59K, 55K, 53K, and 27K as determined by immunoblot analyses. These results demonstrate that the structural gene coding for type 5 streptococcal M protein which was inserted into lambda DNA expresses immunoprotective epitopes, some of which are shared with human heart tissue.

Original languageEnglish (US)
Pages (from-to)198-203
Number of pages6
JournalInfection and Immunity
Volume48
Issue number1
StatePublished - Jun 26 1985

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Epitopes
Escherichia coli
Proteins
Bacteriophages
Streptococcus
Immune Sera
Molecular Weight
Bacteriophage lambda
Myeloma Proteins
Antigens
Precipitins
Peptides
streptococcal M protein
Antibodies
Immunodiffusion
Streptococcus pyogenes
Sodium Dodecyl Sulfate
Genes
Digestion
Membrane Proteins

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

Cite this

Expression of protective and cardiac tissue cross-reactive epitopes of type 5 streptococcal M protein in Escherichia coli. / Poirier, T. P.; Kehoe, M. A.; Dale, James; Timmis, K. N.; Beachey, E. H.

In: Infection and Immunity, Vol. 48, No. 1, 26.06.1985, p. 198-203.

Research output: Contribution to journalArticle

Poirier, T. P. ; Kehoe, M. A. ; Dale, James ; Timmis, K. N. ; Beachey, E. H. / Expression of protective and cardiac tissue cross-reactive epitopes of type 5 streptococcal M protein in Escherichia coli. In: Infection and Immunity. 1985 ; Vol. 48, No. 1. pp. 198-203.
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