Skip to main content
SpringerLink
Log in

Immunological and Biochemical Characterization of Membrane Proteins

  • Protocol
Mycoplasma Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 104))

Abstract

Membrane proteins of mycoplasmas assume particular significance in investigations of the pathogenesis of disease. They are involved in adhesion to, and probably in other interactions with, the cells of the target tissues of the host (1–5). They are frequently significant antigens in the immune response (4–7), and in some cases have been observed to undergo high-frequency size and/or phase variation (8–13). In addition, many membrane proteins are also important in metabolic pathways, including transmembrane transport (14). The significance of future research on membrane proteins of mycoplasmas has been recently highlighted by the completion of the genomic sequences of Mycoplasma genitalium and Mycoplasma pneumoniae (15,16). In both species, the products of a number of open reading frames could not be assigned a function by comparison to known prokaryotic genes. The vast majority of these unassigned products were putative membrane proteins.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Bredt, W., Feldner, J., and Kahane, I. (1981) Adherence of mycoplasmas to cells and inert surfaces: phenomena, experimental models and possible mechanisms. Isr. J. Med. Sci. 17, 586–588.

    PubMed  CAS  Google Scholar 

  2. Kahane, I., Granek, J., and Reisch-Saada, A. (1984) The adhesins of Mycoplasma gallisepticum and M. pneumoniae. Ann. Microbiol. (Inst. Pasteur) 135A, 25–32.

    Article  CAS  Google Scholar 

  3. Dallo, S. F., Su, C. J., Horton, J. R., and Baseman, J. B. (1988) Identification of P1 gene domain containing epitope(s) mediating Mycoplasma pneumoniae cytoadherence. J. Exp. Med. 167, 718–723.

    Article  PubMed  CAS  Google Scholar 

  4. Forsyth, M. H., Tourtellotte, M. E., and Geary, S. J. (1992) Localization of an immunodominant 64 kDa lipoprotein (LP 64) in the membrane of Mycoplasma gallisepticum and its role in cytadherence. Mol. Microbiol. 6, 2099–2106.

    Article  PubMed  CAS  Google Scholar 

  5. Markham, P. F., Glew, M. D., Brandon, M. R., Walker, I. D., and Whithear, K. G. (1992) Characterization of a major hemagglutinin protein from Mycoplasma gallisepticum. Infect. Immun. 60, 3885–3891.

    PubMed  CAS  Google Scholar 

  6. Futo, S., Seto, Y., Okada, M., Sato, S., Suzuki, T., Kawai, K., et al. (1995) Recombinant 46-kilodalton surface antigen (P46) of Mycoplasma hyopneumoniae expressed in Escherichia coli can be used for early specific diagnosis of mycoplasmal pneumonia of swine by enzyme-linked immunosorbent assay. J. Clin. Microbiol. 33, 680–683.

    PubMed  CAS  Google Scholar 

  7. Jan, G., Fontenelle, C., Le, H. M., and Wroblewski, H. (1995) Acylation and immunological properties of Mycoplasma gallisepticum membrane proteins. Res. Microbiol. 146, 739–750.

    Article  PubMed  CAS  Google Scholar 

  8. Boyer, M. J. and Wise, K. S. (1989) Lipid-modified surface protein antigens expressing size variation within the species Mycoplasma hyorhinis. Infect. Immun. 57, 245–254.

    PubMed  CAS  Google Scholar 

  9. Talkington, D. F., Fallon, M. T., Watson, H. L., Thorp, R. K., and Cassell, G. H. (1989) Mycoplasma pulmonis V-1 surface protein variation: occurrence in vivo and association with lung lesions. Microb. Patho. 7, 429–436.

    Article  CAS  Google Scholar 

  10. Rosengarten, R. and Wise, K. S. (1990) Phenotypic switching in mycoplasmas: phase variation of diverse surface lipoproteins. Science 247, 315–318.

    Article  PubMed  CAS  Google Scholar 

  11. Rosengarten, R. and Wise, K. S. (1991) The Vlp system of Mycoplasma hyorhinis: combinatorial expression of distinct size variant lipoproteins generating high-frequency surface antigenic variation. J. Bacteriol. 173, 4782–4793.

    PubMed  CAS  Google Scholar 

  12. Behrens, A., Heller, M., Kirchhoff, H., Yogev, D., and Rosengarten, R. (1994) A family of phase-and size-variant membrane surface lipoprotein antigens (Vsps) of Mycoplasma bovis. Infect. Immun. 62, 5075–5084.

    PubMed  CAS  Google Scholar 

  13. Droesse, M., Tangen, G., Gummelt, I., Kirchhoff, H., Washburn, L. R., and Rosengarten, R. (1995) Major membrane proteins and lipoproteins as highly variable immunogenic surface components and strain-specific antigenic markers of Mycoplasma arthritidis. Microbiology 141, 3207–3219.

    Article  PubMed  CAS  Google Scholar 

  14. Gilson, E., Alloing, G., Schmidt, T., Claverys, J. P., Dudler, R., and Hofnung, M. (1988) Evidence for high affinity binding-protein dependent transport systems in gram-positive bacteria and in Mycoplasma. EMBO J. 7, 3971–3974.

    PubMed  CAS  Google Scholar 

  15. Fraser, C. M., Gocayne, J. D., White, O., Adams, M. D., Clayton, R. A., Fleischmann, R. D., et al. (1995) The minimal gene complement of Mycoplasma genitalium. Science 270, 397–403.

    Article  PubMed  CAS  Google Scholar 

  16. Himmelreich, R., Hilbert, H., Plagens, H., Pirkl, E., Li, B.-C., and Herrmann, R. (1996) Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae. Nucleic Acids Res. 24, 4420–4449.

    Article  PubMed  CAS  Google Scholar 

  17. Brenner, C., Jan, G., Chevalier, Y., and Wroblewski, H. (1995) Evaluation of the efficacy of zwitterionic dodecyl carboxybetaine surfactants for the extraction and the separation of mycoplasma membrane protein antigens. Anal. Biochem. 224, 515–523.

    Article  PubMed  CAS  Google Scholar 

  18. Bordier, C. (1981) Phase separation of integral membrane proteins in Triton X-114 solution. J. Biol. Chem. 256, 1604–1607.

    PubMed  CAS  Google Scholar 

  19. Jan, G., Fontenelle, C., Verrier, F., Le, H. M., and Wroblewski, H. (1996) Selective acylation of plasma membrane proteins of Mycoplasma mycoides subsp. mycoides SC, the contagious bovine pleuropneumonia agent. Curr. Microbiol. 32, 38–42.

    Article  PubMed  CAS  Google Scholar 

  20. Beall, J. A. and Mitchell, G. F. (1986) Identification of a particular antigen from a parasite cDNA library using antibodies affinity purified from selected portions of Western blots. J. Immunol. Methods 86, 217–223.

    Article  PubMed  CAS  Google Scholar 

  21. Bairoch, A. (1993) The PROSITE dictionary of sites and patterns in proteins, its current status. Nucleic Acids Res. 21, 3097–3103.

    Article  PubMed  CAS  Google Scholar 

  22. Bairoch, A. and Bucher, P. (1994) PROSITE: recent developments. Nucleic Acids Res. 22, 3583–9.

    Article  PubMed  CAS  Google Scholar 

  23. Yogev, D., Rosengarten, R., Watson, M. R., and Wise, K. S. (1991) Molecular basis of Mycoplasma surface antigenic variation: a novel set of divergent genes undergo spontaneous mutation of periodic coding regions and 5′ regulatory sequences. EMBO J. 10, 4069–4079.

    PubMed  CAS  Google Scholar 

  24. Wise, K. S., Kim, M. F., Theiss, P. M., and Lo, S. C. (1993) A family of strain-variant surface lipoproteins of Mycoplasma fermentans. Infect. Immun. 61, 3327–3333.

    PubMed  CAS  Google Scholar 

  25. Cleavinger, C. M., Kim, M. F., and Wise, K. S. (1994) Processing and surface presentation of the Mycoplasma hyorhinis variant lipoprotein VlpC. J. Bacteriol. 176, 2463–2467.

    PubMed  CAS  Google Scholar 

  26. Markham, P. F., Glew, M. D., Sykes, J. E., Bowden, T. R., Pollocks, T. D., Browning, G. F., et al. (1994) The organisation of the multigene family which encodes the major cell surface protein, pMGA, of Mycoplasma gallisepticum. FEBS Lett. 352, 347–352.

    Article  PubMed  CAS  Google Scholar 

  27. Ferris, S., Watson, H. L., Neyrolles, O., Montagnier, L., and Blanchard, A. (1995) Characterization of a major Mycoplasma penetrans lipoprotein and of its gene. FEMS Microbiol. Lett. 130, 313–319.

    Article  PubMed  CAS  Google Scholar 

  28. Futo, S., Seto, Y., Mitsuse, S., Mori, Y., Suzuki, T., and Kawai, K. (1995) Molecular cloning of a 46-kilodalton surface antigen (P46) gene from Mycoplasma hyopneumoniae: direct evidence of CGG codon usage for arginine. J. Bacteriol. 177, 1915–1917.

    PubMed  CAS  Google Scholar 

  29. Foissac, X., Saillard, C., Gandar, J., Zreik, L., and Bove, J. M. (1996) Spiralin polymorphism in strains of Spiroplasma citri is not due to differences in post-translational palmitoylation. J. Bacteriol. 178, 2934–2940.

    PubMed  CAS  Google Scholar 

  30. Lysnyansky, I., Rosengarten, R., and Yogev, D. (1996) Phenotypic switching of variable surface lipoproteins in Mycoplasma bovis involves high-frequency chromosomal rearrangements. J. Bacteriol. 178, 5395–5401.

    PubMed  CAS  Google Scholar 

  31. Simmons, W. L., Zuhua, C., Glass, J. I., Simecka, J. W., Cassell, G. H., and Watson, H. L. (1996) Sequence analysis of the chromosomal region around and within the V-1-encoding gene of Mycoplasma pulmonis: evidence for DNA inversion as a mechanism for V-1 variation. Infect. Immun. 64, 472–479.

    PubMed  CAS  Google Scholar 

  32. Zhang, Q. and Wise, K. S. (1996) Molecular basis of size and antigenic variation of a Mycoplasma hominis adhesin encoded by divergent vaa genes. Infect. Immun. 64, 2737–3744.

    PubMed  CAS  Google Scholar 

  33. Djordjevic, S. P., Eamens, G. J., Scarman, A. L., and Chin, J. C. (1996) Characterisation of Mycoplasma hyopneumoniae membrane proteins and identification of discrete antigens which significantly reduce lung pathology in vaccinated pigs challenged with a virulent strain of M. hyopneumoniae. IOM Lett. 4, 282–283.

    Google Scholar 

  34. Wessel, D. and Flugge, U. I. (1984) A method for quantitative recovery of protein in dilute solutions in the presence of detergents and lipids. Anal. Biochem. 138, 141–143.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Veterinary Preclinical Centre, University of Melbourne, Parkville, Victoria, Australia

    Michael F. Duffy, Amir H. Noormohammadi, Nina Baseggio, Glenn F. Browning & Philip F. Markham

Authors
  1. Michael F. Duffy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amir H. Noormohammadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nina Baseggio
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Glenn F. Browning
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Philip F. Markham
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. King’s College, London, England

    Roger Miles

  2. Central Veterinary Laboratory, Surrey, England

    Robin Nicholas

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Duffy, M.F., Noormohammadi, A.H., Baseggio, N., Browning, G.F., Markham, P.F. (1998). Immunological and Biochemical Characterization of Membrane Proteins. In: Miles, R., Nicholas, R. (eds) Mycoplasma Protocols. Methods in Molecular Biology™, vol 104. Humana Press. https://doi.org/10.1385/0-89603-525-5:279

Download citation

  • DOI: https://doi.org/10.1385/0-89603-525-5:279

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-525-6

  • Online ISBN: 978-1-59259-269-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation