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Volume 144, Issue 4

Affinity purification and characterization of a fibrinogen-binding protein complex which protects mice against lethal challenge with Streptococcus Free

Abstract

Cell-wall-associated proteins from subsp. , the causative agent of strangles, were analysed with a view to identifying a potential protective antigen. Preparations of these proteins, isolated from mutanolysin extracts of cell walls, were shown to contain one major high- protein species (apparent 220000 and 550000 when analysed by SDS-PAGE and gel-filtration chromatography, respectively). The high- protein bound horse fibrinogen and was purified under non-denaturing conditions using fibrinogen affinity chromatography. The fibrinogen-binding protein (FgBP) reacted with serum taken from horses recovering from strangles and protected mice against lethal challenge from subsp. . The sequence of the corresponding gene () was determined and shown to encode a mature protein ( 54597) with predicted coiled-coil structure. An FgBP truncate, lacking the C-terminal cell wall/membrane anchor domain, was overexpressed in and purified from and was shown to behave in an analogous fashion to the wild-type product in terms of estimation, fibrinogen binding and seroreactivity.

  • Received:
  • Accepted:
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Keyword(s): equi , fibrinogen-binding protein , protective antigen and Streptococcus equisubsp
© Society for General Microbiology 1998
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1998-04-01
2025-04-27
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References

  1. AlonsoDeVelasco E., Verheul A. F. M., Verhoef J., Snippe H. (1995); Streptococcus pneumoniae-, virulence factors, pathogenesis, and vaccines.. Microbiological Reviews, 59:(4),591–603 [View Article]
     [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. et al. (1990); Basic local alignment search tool, Synthetic peptide vaccine against mucosal colonization by group A streptococci. I. Protection against a heterologous M serotype with shared C repeat region epitopes.. Journal of Molecular Biology, 215:(3),403–110 [View Article]
     [Google Scholar]
  3. Boschwitz J. S., Timoney J. F. (1994); Characterization of the antiphagocytic activity of equine fibrinogen for Streptococcus equi subsp. equi.. Microb Pathog, 17:(2),121–129 [View Article]
     [Google Scholar]
  4. M. H., Timoney J. F. (1991); A comparison of different methods of extraction of the M-protein from Streptococcus equi. Cornell Veterinarian, 8125–36
     [Google Scholar]
  5. Bronze M. S., McKinsey D. S., Beachey E. H., Dale J. B. (1988); Protective immunity evoked by locally administered group A streptococcal vaccines in mice. J Immunol, 1412767–2770
     [Google Scholar]
  6. Bronze M. S., Courtney H. S., Dale J. B. (1992); Epitopes of group A streptococcal M protein that evoke cross-protective local immune responses.. Journal of Immunology, 148:888–893
     [Google Scholar]
  7. Caffrey P., McVeigh T., Owen P. (1988) Western immuno- blotting.. Edited by Owen P., Foster C T. J. In Immunochemical and Molecular Genetic Analysis of Bacterial Pathogens Amsterdam:: Elsevier;255–266
     [Google Scholar]
  8. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. (1977); Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis.. Journal of Biological Chemistry, 252:(3),1102–1106 [View Article]
     [Google Scholar]
  9. D\\'Alessandri R., Plotkin G., Kluge R. M., Wittner M. K., Fox E. N. et al. (1978); Protective studies with group A streptococcal M protein vaccine. III. Challenge of volunteers after systemic or intranasal immunization with type 3 or type 12 group A Streptococcus.. Journal of Infectious Diseases, 138:(6),712–718 [View Article]
     [Google Scholar]
  10. Doherty H., Yamada H., Caffrey P., Owen P. (1986); Identification, immunochemical characterization, and purification of a major lipoprotein antigen associated with the inner (cytoplasmic) membrane of Escherichia coli. Journal of Bacteriology, 1661072–1082
     [Google Scholar]
  11. Dulley J. R., Grieve P. A. (1975); A simple technique for eliminating interference by detergents in the Lowry method of protein determination.. Analytical Biochemistry, 64:(1),136–141 [View Article]
     [Google Scholar]
  12. Fahnestock S. R., Alexander P., Naµle J., Filpula D. (1986); Gene for an immunoµlobulin-binding protein from a group G streptococcus. Journal of Bacteriology, 167870–880
     [Google Scholar]
  13. Feliciello I., Chinali G. (1993); A modified alkaline lysis method for the preparation of highly purified plasmid DNA from Escherichia coli.. Analytical Biochemistry, 212:(2),394–401 [View Article]
     [Google Scholar]
  14. Fischetti V. A. (1991); Streptococcal M-protein.. Scientific American, 264:32–39
     [Google Scholar]
  15. Gal£n J. E., Timoney J. F. (1985); Mucosal nasopharyngeal immune responses of horses to protein antigens of Streptococcus equi.. Infection and Immunity, 47:(3),623–628 [View Article]
     [Google Scholar]
  16. Galcin J. E., Timoney J. F. (1987); Molecular analysis of the M protein of Streptococcus equi and cloning and expression of the M protein gene in Escherichia coli.. Infection and Immunity, 55:(12),3181–3187 [View Article]
     [Google Scholar]
  17. Gal£n J. E., Timoney J. F. (1988); Immunologic and genetic comparison of Streptococcus equi isolates from the United States and Europe.. Journal of Clinical Microbiology, 26:(6),1142–1146 [View Article]
     [Google Scholar]
  18. Galán J. E., Timoney J. F., Curtiss R. Ill (1988); Expression and localization of the Streptococcus equi M protein in Escherichia coli and Salmonella typhimurium.. Equine Infect Dis, 5:34–40
     [Google Scholar]
  19. Gamier J., Osguthorpe D. J., Robson B. (1978); Analysis of the accuracy and implications of simple methods for predicting the secondary structure of µlobular proteins.. ] Mol Biol, 120:(1),97–120 [View Article]
     [Google Scholar]
  20. Goward C. R., Scawen M. D., Murphy J. P., Atkinson T. (1993); Molecular evolution of bacterial cell-surface proteins. Trends in Biochemical Sciences, 18136–140
     [Google Scholar]
  21. Harlow E., Lane D. (1988) Labeling antibodies.. In Antibodies: a Laboratory Manual Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;319–358
     [Google Scholar]
  22. Hartford O., Francois P., Vaudaux P., Foster T. J. (1997); The dipeptide repeat region of the fibrinogen-binding protein (clumping factor) is required for functional expression of the fibrinogen-binding domain on the Staphylococcus aureus cell surface. Molecular Microbiology, 251065–1076
     [Google Scholar]
  23. Hasty D. L., Ofek I., Courtney H. S., Doyle R. J. (1992); Multiple adhesins of streptococci. Infection and Immunity, 602147–2152
     [Google Scholar]
  24. von Heijne G. (1986); A new method for predicting signal sequence cleavage sites.. Nucleic Acids Research, 14:(11),4683–4690 [View Article]
     [Google Scholar]
  25. Hollingshead S. K., Fischetti V. A., Scott J. R. (1986); Complete nucleotide sequence of type 6M protein of the group A Streptococcus : repetitive structure and membrane anchor.. Journal of Biological Chemistry, 261:(4),1677–1686 [View Article]
     [Google Scholar]
  26. Jean-François M. J. B., Poskitt D. C., Turnbull S. J., MacDonald L. M., Yasmeen D. (1991); Protection against Streptococcus equi infection by monoclonal antibodies against an M-like protein.. Journal of General Microbiology, 1372125–2133
     [Google Scholar]
  27. Jonsson H., Lindmark H., Guss B. (1995); A protein G-related cell surface protein in Streptococcus zooepidemicus.. Infection and Immunity, 63:(8),2968–2975 [View Article]
     [Google Scholar]
  28. Jdnsson K., Signets C., Muller H.-P., Lindberg M. (1991); Two different genes encode fibronectin binding proteins in Staphylococcus aureus: the complete nucleotide sequence and characterization of the second gene. Eur ] Biochem, 2021041–1048
     [Google Scholar]
  29. Jorm L. R., Love D. N., Bailey G. D., McKay G. M., Briscoe D. A. (1994); Genetic structure of populations of ^-haemolytic Lancefield group C streptococci from horses and their association with disease.. Research in Veterinary Science, 57:(3),292–299 [View Article]
     [Google Scholar]
  30. Katz J., Harmon G. G., Buckner G. P., Richardson G. J., , Russell M.W., Michalek S.M. (1993); Protective salivary immunoµlobulin A responses against Streptococcus mutans infection after intranasal immunization with S. mutans antigen I/II coupled to the B subunit of cholera toxin.. Infection and Immunity, 61(5),1964–1971 [View Article]
     [Google Scholar]
  31. Kehoe M. A. (1994) Cell-wall-associated proteins in Grampositive bacteria.. Edited by Ghuysen J.-M., Hakenbeck R. New Comprehensive Biochemistry vol. 27 Amsterdam:: Bacterial Cell Wall;217—261
     [Google Scholar]
  32. Laemmli U. K. (1970); Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature, 227:(5259),680–685 [View Article]
     [Google Scholar]
  33. Lech K., Brent R. (1997) Media preparation and bacteriological tools.. Edited by Ausubel F. M. In Current Protocols in Molecular Biology New York:: Wiley;1.1.1–1.1.6
     [Google Scholar]
  34. Lindmark H., Jacobsson K., Frykberg L., Guss B. (1996); Fibronectin-binding protein of Streptococcus equi subsp. zooepidemicus. Infection and Immunity, 643993–3999
     [Google Scholar]
  35. Lupas A. (1996); Prediction and analysis of coiled-coil structures.. Methods in Enzymology, Vol 46: Research on Nitrification and Related Processes, Pt B, 266:513–525
     [Google Scholar]
  36. Lupas A., Van Dyke M., Stock J. (1991); Predicting coiled coils from protein sequences.. Science, 252:(5009),1162–1164 [View Article]
     [Google Scholar]
  37. McVeigh T., Caffrey P., Owen P. (1988) Silver staining of proteins and lipopolysaccharides in SDS-polyacrylamide gels.. Edited by Owen P., Foster C T. J. In Immunochemical and Molecular Genetic Analysis of Bacterial Pathogens Amsterdam:: Elsevier;267–279
     [Google Scholar]
  38. Maniatis T., Fritsch E. F., Sambrook J. (1982) Extraction of bacteriophage X DNA. . In Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory;
     [Google Scholar]
  39. Mosher D. F., Schad P. E., Vann J. M. (1980); Cross-linking of collagen and fibronectin by factor XHIa. Localization of participating µlutaminyl residues to a tryptic fragment of fibronectin.. Journal of Biological Chemistry, 255:(3),1181–1188 [View Article]
     [Google Scholar]
  40. Newell D. G., McBride B. W., Clark S. A. (1988) Enzyme-linked immunosorbent assay for soluble antigens. . In Making Mono- clonals: a Practical Beginners Guide to the Production and Characterization of Monoclonal Antibodies against Bacteria and Viruses Colindale, UK:: Public Health Laboratory Service;37–39
     [Google Scholar]
  41. Nilson B. H. K., Frick l-M., Akesson P., Fors6n S., Bj&rck L. et al. (1995); Structure and stability of protein H and the Ml protein from Streptococcus pyogenes. Implications for other surface proteins of Gram-positive bacteria.. Biochemistry, 34:(41),13688–13698 [View Article]
     [Google Scholar]
  42. Okahashi N., Sasakawa C., Yoshikawa M., Hamada S., Koga T. (1989); Molecular characterization of a surface protein antigen from serotype c Streptococcus mutans, implicated in dental caries.. Molecular Microbiology, 3:(5),673–678 [View Article]
     [Google Scholar]
  43. O'Reilly M., O'Toole P. W., Foster T. J. (1988) Screening lambda libraries and detection of recombinants.. Edited by Owen P., Foster T. J. In Immunochemical and Molecular Genetic Analysis of Bacterial Pathogens Amsterdam:: Elsevier;187–197
     [Google Scholar]
  44. Owen P. (1985) Crossed immunoelectrophoresis in the study of outer membrane antigens.. In: Edited by Korhonen T. K., Dawes E. A., Makela P. H. Enterobacterial Surface Antigens: Methods for Molecular Characterization Amsterdam:: Elsevier;207–242
     [Google Scholar]
  45. Owen P. (1986); An improved procedure for polypeptide analysis of radiolabeled antigens resolved by crossed immunoelectrophoresis and its application to the study of inner and outer membranes of Escherichia coli.. Electrophoresis, 7:(1),19–28 [View Article]
     [Google Scholar]
  46. Patti J. M., Allen B. L., McGavin M. J., Hdandk M. (1994); MSCRAMM-mediated adherence of microorganisms to host tissues. Annual Review of Microbiology, 45585–617
     [Google Scholar]
  47. Polly S. M., Waldman R. H., High P., Wittner M. K., Dorfman A. et al. (1975); Protective studies with a group A streptococcal M protein vaccine. II. Challenge of volunteers after local immunization in the upper respiratory tract.. Journal of Infectious Diseases, 131:(3),217–224 [View Article]
     [Google Scholar]
  48. Schennings T., Heimdahi A., Coster K., Flock J.-l. (1993); Immunization with fibronectin binding protein from Staphylococcus aureus protects against experimental endocarditis in rats. Microb Pathog, 15227–236
     [Google Scholar]
  49. Talay S. R., Grammel M. P., Chhatwal G. S. (1996); Structure of a group C streptococcal protein that binds to fibrinogen, albumin and immunoµlobulin G via overlapping modules.. Biochemical Journal, 315:(2),577–582 [View Article]
     [Google Scholar]
  50. Timoney J. F. (1988); Protecting against ‘stranµles’: a contemporary view.. Equine Veterinary Journal, 20:(6),392–394 [View Article]
     [Google Scholar]
  51. Timoney J. F. (1993); Stranµles.. Vet Clin North Am Equine Pract, 9:(2),365–374 [View Article]
     [Google Scholar]
  52. Timoney J. F., Mukhtar M. M. (1993); The protective M proteins of the equine group C streptococci.. Veterinary Microbiology, 37:(3–4),389–395 [View Article]
     [Google Scholar]
  53. Timoney J. F., Trachman J. (1985); Immunologically reactive proteins of Streptococcus equi.. Infection and Immunity, 48:(1),29–34 [View Article]
     [Google Scholar]
  54. Timoney J. F., Mukhtar M., Galdn J., Ding J. (1991),; M proteins of the equine group C streptococci. Edited by Dunny G. M., Cleary P. P., McKay L. L. Genetics and Molecular Biology of Streptococci, Lactococci, and Enterococci Washington, DC:: American Society for Microbiology;160–164
     [Google Scholar]
  55. Timoney J. F., Walker J., Zhou M., Ding J. (1995); Cloning and sequence analysis of a protective M-like protein gene from Streptococcus equi subsp. zooepidemicus.. Infection and Immunity, 63:(4),1440–1445 [View Article]
     [Google Scholar]
  56. Timoney J. F., Artiushin S. C. (1997); 8i Boschwitz, Comparison of the sequences and functions of Streptococcus equi M- like proteins SeM and SzPSe.. J., 5:3600–3605
     [Google Scholar]
  57. Wilson K. (1997) Preparation of genomic DNA from bacteria.. Edited by Ausubel F. M. Current Protocols in Molecular Biology New York:: Wiley;pp 2.4.1–2.4.5
     [Google Scholar]
  58. Wolf E., Kim P. S., Berger B. (1997); MultiCoil: a program for predicting two- and three-stranded coiled-coils.. Protein Science, 6:(6),1179–1189 [View Article]
     [Google Scholar]
  59. Yelle M. T. (1987); Clinical aspects of Streptococcus equi infection.. Equine Veterinary Journal, 19:(2),158–162 [View Article]
     [Google Scholar]
  60. Yu C.-E., Ferretti J. J. (1989); Molecular epidemiologic analysis of the type A streptococcal exotoxin (erythrogenic toxin) gene (speA) in clinical Streptococcus pyogenes strains.. Infection and Immunity, 57:(12),3715–3719 [View Article]
     [Google Scholar]
/content/journal/micro/10.1099/00221287-144-4-993
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Affinity purification and characterization of a fibrinogen-binding protein complex which protects mice against lethal challenge with Streptococcus equi subsp. equi
Microbiology 144, 993 (1998); https://doi.org/10.1099/00221287-144-4-993
/content/journal/micro/10.1099/00221287-144-4-993
/content/journal/micro/10.1099/00221287-144-4-993
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