Molecular cloning, purification and immunological responses of recombinants GroEL and DnaK from Streptococcus pyogenes.
To better understand the roles of heat shock proteins in streptococcal diseases, the groEL and dnaK genes from Streptococcus pyogenes were cloned and their products (GroEL and DnaK) and derivatives (F2GroEL, F3GroEL and C1DnaK) purified as His-tagged fusion proteins. Western blot analysis of the purified proteins with sera from individuals with streptococcal diseases demonstrated that 29 out of 36 sera tested were reactive with GroEL and eight recognized DnaK. Rabbit antiserum against myosin recognized both GroEL and DnaK. Antibodies raised against purified F2GroEL and DnaK reacted with myosin in the ELISA but not in a Western immunoblot. These data indicate that the S. pyogenes GroEL and DnaK may be important immunogens during streptococcal infections. Furthermore, we provide evidence of an immunogenic relatedness of the GroEL and DnaK proteins with myosin that could play a role in the pathogenesis of streptococcal non-suppurative sequelae.
Lemos
J A
JA
Instituto de Microbiologia, CCS, Universidade Federal do Rio de Janeiro, Ilha do Fundão, Rio de Janeiro, Brazil.
Burne
R A
RA
Castro
A C
AC
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D015153:Blotting, Western; D018834:Chaperonin 60 / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D006801:Humans; D009218:Myosins / Q000276:immunology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013290:Streptococcal Infections / Q000276:immunology; D013297:Streptococcus pyogenes / Q000737:chemistry / Q000235:genetics*
D000907:Antibodies, Bacterial; D018834:Chaperonin 60; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; C063842:dnaK protein, E coli; D009218:Myosins