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. 1996 Jul;64(7):2787–2793. doi: 10.1128/iai.64.7.2787-2793.1996

Identification of an immunoglobulin A binding motif located in the beta-antigen of the c protein complex of group B streptococci.

P G Jerlström 1, S R Talay 1, P Valentin-Weigand 1, K N Timmis 1, G S Chhatwal 1
PMCID: PMC174140  PMID: 8698509

Abstract

The beta-antigen of the c protein complex of group B streptococci contains two immunoglobulin A (IgA)-binding domains called A and B. A 73-amino-acid segment in domain A is responsible for most of the IgA-binding activity. To identify the IgA binding motif, the 73-amino-acid domain was divided into 60 14-amino-acid overlapping peptides spot synthesized onto a cellulose membrane. A 20-residue putative antigenic epitope was identified and expressed as a fusion protein. The fusion protein was purified by fast protein liquid chromatography and used to raise rabbit antiserum. By use of a membrane with spot-synthesized peptide amino acids of decreasing length (from 14 to 6 amino acids), the major antigenic epitope recognized by the anti-fusion protein antibodies was mapped to motif MLKKIE. Anti-fusion protein antibodies inhibited the binding of IgA to group B streptococci. This inhibition could be blocked by the peptide containing the motif MLKKIE. These results indicate that the motif MLKKIE is located in the IgA-binding site. The IgA-binding domain of beta-antigen from three group B streptococcal strains reacted with the anti-fusion protein antibodies, and their coding sequences gave positive signals in Southern hybridization. The sequences of beta-antigen from these strains were amplified by PCR, and sequence analysis showed them to be identical. The results indicate that the motif MLKKIE is required for IgA binding and is present in different group B streptococcal strains.

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Selected References

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  1. Allison L. A., Moyle M., Shales M., Ingles C. J. Extensive homology among the largest subunits of eukaryotic and prokaryotic RNA polymerases. Cell. 1985 Sep;42(2):599–610. doi: 10.1016/0092-8674(85)90117-5. [DOI] [PubMed] [Google Scholar]
  2. Aucouturier P., Mihaesco E., Mihaesco C., Preud'homme J. L. Characterization of jacalin, the human IgA and IgD binding lectin from jackfruit. Mol Immunol. 1987 May;24(5):503–511. doi: 10.1016/0161-5890(87)90025-3. [DOI] [PubMed] [Google Scholar]
  3. Baker C. J. Vaccine prevention of group B streptococcal disease. Pediatr Ann. 1993 Dec;22(12):711–714. doi: 10.3928/0090-4481-19931201-05. [DOI] [PubMed] [Google Scholar]
  4. Bevanger L. Ibc proteins as serotype markers of group B streptococci. Acta Pathol Microbiol Immunol Scand B. 1983 Aug;91(4):231–234. doi: 10.1111/j.1699-0463.1983.tb00038.x. [DOI] [PubMed] [Google Scholar]
  5. Bevanger L., Naess A. I. Mouse-protective antibodies against the Ibc proteins of group B streptococci. Acta Pathol Microbiol Immunol Scand B. 1985 Apr;93(2):121–124. doi: 10.1111/j.1699-0463.1985.tb02862.x. [DOI] [PubMed] [Google Scholar]
  6. Biewenga J., Steneker I., Hameleers D. M. Effect of serum albumin on the recovery of human IgA1 from immobilized jacalin. J Immunol Methods. 1988 Dec 9;115(2):199–207. doi: 10.1016/0022-1759(88)90289-x. [DOI] [PubMed] [Google Scholar]
  7. Brady L. J., Boyle M. D. Identification of non-immunoglobulin A-Fc-binding forms and low-molecular-weight secreted forms of the group B streptococcal beta antigen. Infect Immun. 1989 May;57(5):1573–1581. doi: 10.1128/iai.57.5.1573-1581.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brady L. J., Daphtary U. D., Ayoub E. M., Boyle M. D. Two novel antigens associated with group B streptococci identified by a rapid two-stage radioimmunoassay. J Infect Dis. 1988 Nov;158(5):965–972. doi: 10.1093/infdis/158.5.965. [DOI] [PubMed] [Google Scholar]
  9. Chen E. Y., Seeburg P. H. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. doi: 10.1089/dna.1985.4.165. [DOI] [PubMed] [Google Scholar]
  10. Chhatwal G. S., Albohn G., Blobel H. Novel complex formed between a nonproteolytic cell wall protein of group A streptococci and alpha 2-macroglobulin. J Bacteriol. 1987 Aug;169(8):3691–3695. doi: 10.1128/jb.169.8.3691-3695.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chhatwal G. S., Preissner K. T., Müller-Berghaus G., Blobel H. Specific binding of the human S protein (vitronectin) to streptococci, Staphylococcus aureus, and Escherichia coli. Infect Immun. 1987 Aug;55(8):1878–1883. doi: 10.1128/iai.55.8.1878-1883.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chun C. S., Brady L. J., Boyle M. D., Dillon H. C., Ayoub E. M. Group B streptococcal C protein-associated antigens: association with neonatal sepsis. J Infect Dis. 1991 Apr;163(4):786–791. doi: 10.1093/infdis/163.4.786. [DOI] [PubMed] [Google Scholar]
  13. Cleary P., Retnoningrum D. Group A streptococcal immunoglobulin-binding proteins: adhesins, molecular mimicry or sensory proteins? Trends Microbiol. 1994 Apr;2(4):131–136. doi: 10.1016/0966-842x(94)90600-9. [DOI] [PubMed] [Google Scholar]
  14. Cleat P. H., Timmis K. N. Cloning and expression in Escherichia coli of the Ibc protein genes of group B streptococci: binding of human immunoglobulin A to the beta antigen. Infect Immun. 1987 May;55(5):1151–1155. doi: 10.1128/iai.55.5.1151-1155.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Delacroix D. L., Malburny G. N., Vaerman J. P. Hepatobiliary transport of plasma IgA in the mouse: contribution to clearance of intravascular IgA. Eur J Immunol. 1985 Sep;15(9):893–899. doi: 10.1002/eji.1830150906. [DOI] [PubMed] [Google Scholar]
  16. Fahnestock S. R., Alexander P., Nagle J., Filpula D. Gene for an immunoglobulin-binding protein from a group G streptococcus. J Bacteriol. 1986 Sep;167(3):870–880. doi: 10.1128/jb.167.3.870-880.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Faulmann E. L., Duvall J. L., Boyle M. D. Protein B: a versatile bacterial Fc-binding protein selective for human IgA. Biotechniques. 1991 Jun;10(6):748–755. [PubMed] [Google Scholar]
  18. Frithz E., Hedén L. O., Lindahl G. Extensive sequence homology between IgA receptor and M proteins in Streptococcus pyogenes. Mol Microbiol. 1989 Aug;3(8):1111–1119. doi: 10.1111/j.1365-2958.1989.tb00261.x. [DOI] [PubMed] [Google Scholar]
  19. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  20. Heard S. R., Mawn J. A. New phenotypic typing scheme for group B streptococci. J Clin Pathol. 1993 Feb;46(2):145–148. doi: 10.1136/jcp.46.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hedén L. O., Frithz E., Lindahl G. Molecular characterization of an IgA receptor from group B streptococci: sequence of the gene, identification of a proline-rich region with unique structure and isolation of N-terminal fragments with IgA-binding capacity. Eur J Immunol. 1991 Jun;21(6):1481–1490. doi: 10.1002/eji.1830210623. [DOI] [PubMed] [Google Scholar]
  22. Jerlström P. G., Chhatwal G. S., Timmis K. N. The IgA-binding beta antigen of the c protein complex of Group B streptococci: sequence determination of its gene and detection of two binding regions. Mol Microbiol. 1991 Apr;5(4):843–849. doi: 10.1111/j.1365-2958.1991.tb00757.x. [DOI] [PubMed] [Google Scholar]
  23. Johnson D. R., Ferrieri P. Group B streptococcal Ibc protein antigen: distribution of two determinants in wild-type strains of common serotypes. J Clin Microbiol. 1984 Apr;19(4):506–510. doi: 10.1128/jcm.19.4.506-510.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kondoh H., Kobayashi K., Hagiwara K., Kajii T. Jacalin, a jackfruit lectin, precipitates IgA1 but not IgA2 subclass on gel diffusion reaction. J Immunol Methods. 1986 Apr 17;88(2):171–173. doi: 10.1016/0022-1759(86)90003-7. [DOI] [PubMed] [Google Scholar]
  25. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  26. Lancefield R. C., McCarty M., Everly W. N. Multiple mouse-protective antibodies directed against group B streptococci. Special reference to antibodies effective against protein antigens. J Exp Med. 1975 Jul 1;142(1):165–179. doi: 10.1084/jem.142.1.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lim D. V., Morales W. J., Walsh A. F., Kazanis D. Reduction of morbidity and mortality rates for neonatal group B streptococcal disease through early diagnosis and chemoprophylaxis. J Clin Microbiol. 1986 Mar;23(3):489–492. doi: 10.1128/jcm.23.3.489-492.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lindahl G., Akerström B., Vaerman J. P., Stenberg L. Characterization of an IgA receptor from group B streptococci: specificity for serum IgA. Eur J Immunol. 1990 Oct;20(10):2241–2247. doi: 10.1002/eji.1830201013. [DOI] [PubMed] [Google Scholar]
  29. Madoff L. C., Paoletti L. C., Tai J. Y., Kasper D. L. Maternal immunization of mice with group B streptococcal type III polysaccharide-beta C protein conjugate elicits protective antibody to multiple serotypes. J Clin Invest. 1994 Jul;94(1):286–292. doi: 10.1172/JCI117319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Michel J. L., Madoff L. C., Kling D. E., Kasper D. L., Ausubel F. M. Cloned alpha and beta C-protein antigens of group B streptococci elicit protective immunity. Infect Immun. 1991 Jun;59(6):2023–2028. doi: 10.1128/iai.59.6.2023-2028.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Musser J. M., Mattingly S. J., Quentin R., Goudeau A., Selander R. K. Identification of a high-virulence clone of type III Streptococcus agalactiae (group B Streptococcus) causing invasive neonatal disease. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4731–4735. doi: 10.1073/pnas.86.12.4731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Olsson A., Eliasson M., Guss B., Nilsson B., Hellman U., Lindberg M., Uhlén M. Structure and evolution of the repetitive gene encoding streptococcal protein G. Eur J Biochem. 1987 Oct 15;168(2):319–324. doi: 10.1111/j.1432-1033.1987.tb13423.x. [DOI] [PubMed] [Google Scholar]
  33. Rubens C. E., Wessels M. R., Heggen L. M., Kasper D. L. Transposon mutagenesis of type III group B Streptococcus: correlation of capsule expression with virulence. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7208–7212. doi: 10.1073/pnas.84.20.7208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Russell-Jones G. J., Gotschlich E. C., Blake M. S. A surface receptor specific for human IgA on group B streptococci possessing the Ibc protein antigen. J Exp Med. 1984 Nov 1;160(5):1467–1475. doi: 10.1084/jem.160.5.1467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Saitoh H., Tomkiel J., Cooke C. A., Ratrie H., 3rd, Maurer M., Rothfield N. F., Earnshaw W. C. CENP-C, an autoantigen in scleroderma, is a component of the human inner kinetochore plate. Cell. 1992 Jul 10;70(1):115–125. doi: 10.1016/0092-8674(92)90538-n. [DOI] [PubMed] [Google Scholar]
  36. Sjöbring U., Björck L., Kastern W. Streptococcal protein G. Gene structure and protein binding properties. J Biol Chem. 1991 Jan 5;266(1):399–405. [PubMed] [Google Scholar]
  37. Stålhammar-Carlemalm M., Stenberg L., Lindahl G. Protein rib: a novel group B streptococcal cell surface protein that confers protective immunity and is expressed by most strains causing invasive infections. J Exp Med. 1993 Jun 1;177(6):1593–1603. doi: 10.1084/jem.177.6.1593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sugino A., Peebles C. L., Kreuzer K. N., Cozzarelli N. R. Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4767–4771. doi: 10.1073/pnas.74.11.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Talay S. R., Ehrenfeld E., Chhatwal G. S., Timmis K. N. Expression of the fibronectin-binding components of Streptococcus pyogenes in Escherichia coli demonstrates that they are proteins. Mol Microbiol. 1991 Jul;5(7):1727–1734. doi: 10.1111/j.1365-2958.1991.tb01921.x. [DOI] [PubMed] [Google Scholar]
  40. Uhlén M., Guss B., Nilsson B., Gatenbeck S., Philipson L., Lindberg M. Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications. J Biol Chem. 1984 Feb 10;259(3):1695–1702. [PubMed] [Google Scholar]
  41. Wilkinson H. W., Eagon R. G. Type-specific antigens of group B type Ic streptococci. Infect Immun. 1971 Nov;4(5):596–604. doi: 10.1128/iai.4.5.596-604.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

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