Abstract
Streptococcus equi subsp. zooepidemicus, a Lancefield group C streptococcus, is a frequently isolated opportunist pathogen from a variety of animal hosts, including the horse. Previous studies have indicated that equine strains carry antigens with characteristics of the antiphagocytic M proteins on the Lancefield groups A and G streptococci. We have cloned a protective M-like protein gene (SzPW60) of an equine strain of S. equi subsp. zooepidemicus W60 and determined its sequence. This gene encodes a protein with a molecular weight of 40,123 which protects mice against subsp. zooepidemicus but not subsp. equi, stimulates antibodies which opsonize subsp. zooepidemicus but not equi, and reacts with antiserum to the protein of the parent strain. The predicted amino acid structure shows significant homology with the carboxy termini of groups A and G M proteins but no other homology. The M-like protein, although showing an extensive region of alpha helix, lacks the A, B, and C repeats found in group A M proteins and has a shorter signal sequence. A proline-rich region upstream from the LPSTGE motif contains 20 repeats of the tetrapeptide PEPK. The presence of this repeat region may account for the slow migration of the M-like protein in sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Full Text
The Full Text of this article is available as a PDF (268.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bessen D. E., Fischetti V. A. Nucleotide sequences of two adjacent M or M-like protein genes of group A streptococci: different RNA transcript levels and identification of a unique immunoglobulin A-binding protein. Infect Immun. 1992 Jan;60(1):124–135. doi: 10.1128/iai.60.1.124-135.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boyle M. D., Hawlitzky J., Raeder R., Podbielski A. Analysis of genes encoding two unique type IIa immunoglobulin G-binding proteins expressed by a single group A streptococcal isolate. Infect Immun. 1994 Apr;62(4):1336–1347. doi: 10.1128/iai.62.4.1336-1347.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clayton C. E., Mowatt M. R. The procyclic acidic repetitive proteins of Trypanosoma brucei. Purification and post-translational modification. J Biol Chem. 1989 Sep 5;264(25):15088–15093. [PubMed] [Google Scholar]
- 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]
- Fischetti V. A. Streptococcal M protein: molecular design and biological behavior. Clin Microbiol Rev. 1989 Jul;2(3):285–314. doi: 10.1128/cmr.2.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Galan J. E., Timoney J. F. Mucosal nasopharyngeal immune responses of horses to protein antigens of Streptococcus equi. Infect Immun. 1985 Mar;47(3):623–628. doi: 10.1128/iai.47.3.623-628.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galán J. E., Timoney J. F. Immunologic and genetic comparison of Streptococcus equi isolates from the United States and Europe. J Clin Microbiol. 1988 Jun;26(6):1142–1146. doi: 10.1128/jcm.26.6.1142-1146.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gomi H., Hozumi T., Hattori S., Tagawa C., Kishimoto F., Björck L. The gene sequence and some properties of protein H. A novel IgG-binding protein. J Immunol. 1990 May 15;144(10):4046–4052. [PubMed] [Google Scholar]
- Hollingshead S. K., Fischetti V. A., Scott J. R. Size variation in group A streptococcal M protein is generated by homologous recombination between intragenic repeats. Mol Gen Genet. 1987 May;207(2-3):196–203. doi: 10.1007/BF00331578. [DOI] [PubMed] [Google Scholar]
- Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
- Kehoe M., Timmis K. N. Cloning and expression in Escherichia coli of the streptolysin O determinant from Streptococcus pyogenes: characterization of the cloned streptolysin O determinant and demonstration of the absence of substantial homology with determinants of other thiol-activated toxins. Infect Immun. 1984 Mar;43(3):804–810. doi: 10.1128/iai.43.3.804-810.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Khandke K. M., Fairwell T., Manjula B. N. Difference in the structural features of streptococcal M proteins from nephritogenic and rheumatogenic serotypes. J Exp Med. 1987 Jul 1;166(1):151–162. doi: 10.1084/jem.166.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LANCEFIELD R. C., PERLMANN G. E. Preparation and properties of type-specific M antigen isolated from a group A, type 1 hemolytic streptococcus. J Exp Med. 1952 Jul;96(1):71–82. doi: 10.1084/jem.96.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller L., Gray L., Beachey E., Kehoe M. Antigenic variation among group A streptococcal M proteins. Nucleotide sequence of the serotype 5 M protein gene and its relationship with genes encoding types 6 and 24 M proteins. J Biol Chem. 1988 Apr 25;263(12):5668–5673. [PubMed] [Google Scholar]
- Moore B. O., Bryans J. T. Antigenic classification of group C animal streptococci. J Am Vet Med Assoc. 1969 Jul 15;155(2):416–421. [PubMed] [Google Scholar]
- Novotný J., Auffray C. A program for prediction of protein secondary structure from nucleotide sequence data: application to histocompatibility antigens. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):243–255. doi: 10.1093/nar/12.1part1.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Piszkiewicz D., Landon M., Smith E. L. Anomalous cleavage of aspartyl-proline peptide bonds during amino acid sequence determinations. Biochem Biophys Res Commun. 1970 Sep 10;40(5):1173–1178. doi: 10.1016/0006-291x(70)90918-6. [DOI] [PubMed] [Google Scholar]
- Pycock J. F., Allen W. E., Morris T. H. Rapid, single-step isolation of equine neutrophils on a discontinuous Percoll density gradient. Res Vet Sci. 1987 May;42(3):411–412. [PubMed] [Google Scholar]
- Sjöbring U. Isolation and molecular characterization of a novel albumin-binding protein from group G streptococci. Infect Immun. 1992 Sep;60(9):3601–3608. doi: 10.1128/iai.60.9.3601-3608.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stewart G. S., Lubinsky-Mink S., Jackson C. G., Cassel A., Kuhn J. pHG165: a pBR322 copy number derivative of pUC8 for cloning and expression. Plasmid. 1986 May;15(3):172–181. doi: 10.1016/0147-619x(86)90035-1. [DOI] [PubMed] [Google Scholar]
- Timoney J. F., Trachman J. Immunologically reactive proteins of Streptococcus equi. Infect Immun. 1985 Apr;48(1):29–34. doi: 10.1128/iai.48.1.29-34.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsonis P. A., Manes T. Rapid phage DNA isolation without the use of enzymes. Biotechniques. 1988 Nov-Dec;6(10):950–951. [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
- von Heijne G., Abrahmsén L. Species-specific variation in signal peptide design. Implications for protein secretion in foreign hosts. FEBS Lett. 1989 Feb 27;244(2):439–446. doi: 10.1016/0014-5793(89)80579-4. [DOI] [PubMed] [Google Scholar]