1887

Abstract

In different, phylogenetically unrelated micro-organisms, glycolytic enzymes play a dual role. In the cytosol they are involved in metabolic reactions whereas the surface-localized fraction of the enzymes contributes to adhesion and virulence. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a typical member of this group of multifunctional proteins. In this study, we characterized the GAPDH of , a common pathogen of the human respiratory mucosa. Full-length GAPDH of was successfully expressed and used to produce a polyclonal antiserum. By immunofluorescence, colony blot and ELISA experiments with different fractions of the proteins, GAPDH was demonstrated to be present in the cytosol and at even higher concentrations at the surface of mycoplasmas. Nevertheless, antibodies against recombinant GAPDH were not detected in sera of immunized animals or of patients with confirmed infection. Recombinant GAPDH bound to different human cell lines in a concentration-dependent manner, and binding was inhibited by specific anti-GAPDH serum. In contrast, this antiserum did not significantly influence the adherence of to HeLa cells. When different human extracellular matrix proteins were tested in Western blot assays, GAPDH bound to fibrinogen. The results showed that the GAPDH of is a member of the family of cytosol-localized glycolytic enzymes, which also occur at the surface of the bacterium, and mediates interactions with the extracellular matrix proteins of the human host. Thus, the surface-exposed fraction of GAPDH may be a factor that contributes to the successful colonization of the human respiratory tract by .

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2011-08-01
2024-03-28
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References

  1. Alvarez R. A., Blaylock M. W., Baseman J. B. ( 2003). Surface localized glyceraldehyde-3-phosphate dehydrogenase of Mycoplasma genitalium binds mucin. Mol Microbiol 48:1417–1425 [View Article][PubMed]
    [Google Scholar]
  2. Balasubramanian S., Kannan T. R., Hart P. J., Baseman J. B. ( 2009). Amino acid changes in elongation factor Tu of Mycoplasma pneumoniae and Mycoplasma genitalium influence fibronectin binding. Infect Immun 77:3533–3541 [View Article][PubMed]
    [Google Scholar]
  3. Barbosa M. S., Báo S. N., Andreotti P. F., de Faria F. P., Felipe M. S., dos Santos Feitosa L., Mendes-Giannini M. J., Soares C. M. ( 2006). Glyceraldehyde-3-phosphate dehydrogenase of Paracoccidioides brasiliensis is a cell surface protein involved in fungal adhesion to extracellular matrix proteins and interaction with cells. Infect Immun 74:382–389 [View Article][PubMed]
    [Google Scholar]
  4. Bendtsen J. D., Kiemer L., Fausbøll A., Brunak S. ( 2005). Non-classical protein secretion in bacteria. BMC Microbiol 5:58 [View Article][PubMed]
    [Google Scholar]
  5. Bergmann S., Rohde M., Hammerschmidt S. ( 2004). Glyceraldehyde-3-phosphate dehydrogenase of Streptococcus pneumoniae is a surface-displayed plasminogen-binding protein. Infect Immun 72:2416–2419 [View Article][PubMed]
    [Google Scholar]
  6. Boël G., Jin H., Pancholi V. ( 2005). Inhibition of cell surface export of group A streptococcal anchorless surface dehydrogenase affects bacterial adherence and antiphagocytic properties. Infect Immun 73:6237–6248 [View Article][PubMed]
    [Google Scholar]
  7. Chumchua V., Pornputtapong N., Thammarongtham C., Meksuriyen D. ( 2008). Homology modeling of Mycoplasma pneumoniae enolase and its molecular interaction with human plasminogen. Bioinformation 3:18–23[PubMed] [CrossRef]
    [Google Scholar]
  8. Dallo S. F., Kannan T. R., Blaylock M. W., Baseman J. B. ( 2002). Elongation factor Tu and E1 β subunit of pyruvate dehydrogenase complex act as fibronectin binding proteins in Mycoplasma pneumoniae . Mol Microbiol 46:1041–1051 [View Article][PubMed]
    [Google Scholar]
  9. Dandekar T., Huynen M., Regula J. T., Ueberle B., Zimmermann C. U., Andrade M. A., Doerks T., Sánchez-Pulido L., Snel B. et al. ( 2000). Re-annotating the Mycoplasma pneumoniae genome sequence: adding value, function and reading frames. Nucleic Acids Res 28:3278–3288 [View Article][PubMed]
    [Google Scholar]
  10. Delgado M. L., Gil M. L., Gozalbo D. ( 2003). Starvation and temperature upshift cause an increase in the enzymatically active cell wall-associated glyceraldehyde-3-phosphate dehydrogenase protein in yeast. FEMS Yeast Res 4:297–303 [View Article][PubMed]
    [Google Scholar]
  11. Dumke R., Catrein I., Pirkil E., Herrmann R., Jacobs E. ( 2003). Subtyping of Mycoplasma pneumoniae isolates based on extended genome sequencing and on expression profiles. Int J Med Microbiol 292:513–525 [View Article][PubMed]
    [Google Scholar]
  12. Dumke R., Von Baum H., Lück P. C., Jacobs E. ( 2010). Subtypes and variants of Mycoplasma pneumoniae: local and temporal changes in Germany 2003-2006 and absence of a correlation between the genotype in the respiratory tract and the occurrence of genotype-specific antibodies in the sera of infected patients. Epidemiol Infect 138:1829–1837 [View Article][PubMed]
    [Google Scholar]
  13. Dutow P., Schmidl S. R., Ridderbusch M., Stülke J. ( 2010). Interactions between glycolytic enzymes of Mycoplasma pneumoniae . J Mol Microbiol Biotechnol 19:134–139 [View Article][PubMed]
    [Google Scholar]
  14. Egea L., Aguilera L., Giménez R., Sorolla M. A., Aguilar J., Badía J., Baldoma L. ( 2007). Role of secreted glyceraldehyde-3-phosphate dehydrogenase in the infection mechanism of enterohemorrhagic and enteropathogenic Escherichia coli: interaction of the extracellular enzyme with human plasminogen and fibrinogen. Int J Biochem Cell Biol 39:1190–1203 [View Article][PubMed]
    [Google Scholar]
  15. Eun B. W., Kim N. H., Choi E. H., Lee H. J. ( 2008). Mycoplasma pneumoniae in Korean children: the epidemiology of pneumonia over an 18-year period. J Infect 56:326–331 [View Article][PubMed]
    [Google Scholar]
  16. Gozalbo D., Gil-Navarro I., Azorín I., Renau-Piqueras J., Martínez J. P., Gil M. L. ( 1998). The cell wall-associated glyceraldehyde-3-phosphate dehydrogenase of Candida albicans is also a fibronectin and laminin binding protein. Infect Immun 66:2052–2059[PubMed]
    [Google Scholar]
  17. Hames C., Halbedel S., Schilling O., Stülke J. ( 2005). Multiple-mutation reaction: a method for simultaneous introduction of multiple mutations into the glpK gene of Mycoplasma pneumoniae . Appl Environ Microbiol 71:4097–4100 [View Article][PubMed]
    [Google Scholar]
  18. Herrmann R., Reiner B. ( 1998). Mycoplasma pneumoniae and Mycoplasma genitalium: a comparison of two closely related bacterial species. Curr Opin Microbiol 1:572–579 [View Article][PubMed]
    [Google Scholar]
  19. Himmelreich R., Hilbert H., Plagens H., Pirkl E., Li B. C., Herrmann R. ( 1996). Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae . Nucleic Acids Res 24:4420–4449 [View Article][PubMed]
    [Google Scholar]
  20. Hoelzle L. E., Hoelzle K., Helbling M., Aupperle H., Schoon H. A., Ritzmann M., Heinritzi K., Felder K. M., Wittenbrink M. M. ( 2007). MSG1, a surface-localised protein of Mycoplasma suis is involved in the adhesion to erythrocytes. Microbes Infect 9:466–474 [View Article][PubMed]
    [Google Scholar]
  21. Kannan T. R., Baseman J. B. ( 2006). ADP-ribosylating and vacuolating cytotoxin of Mycoplasma pneumoniae represents unique virulence determinant among bacterial pathogens. Proc Natl Acad Sci U S A 103:6724–6729 [View Article][PubMed]
    [Google Scholar]
  22. Kinoshita H., Uchida H., Kawai Y., Kawasaki T., Wakahara N., Matsuo H., Watanabe M., Kitazawa H., Ohnuma S. et al. ( 2008). Cell surface Lactobacillus plantarum LA 318 glyceraldehyde-3-phosphate dehydrogenase (GAPDH) adheres to human colonic mucin. J Appl Microbiol 104:1667–1674 [View Article][PubMed]
    [Google Scholar]
  23. Klement E., Talkington D. F., Wasserzug O., Kayouf R., Davidovitch N., Dumke R., Bar-Zeev Y., Ron M., Boxman J. et al. ( 2006). Identification of risk factors for infection in an outbreak of Mycoplasma pneumoniae respiratory tract disease. Clin Infect Dis 43:1239–1245 [View Article][PubMed]
    [Google Scholar]
  24. Krause D. C., Balish M. F. ( 2004). Cellular engineering in a minimal microbe: structure and assembly of the terminal organelle of Mycoplasma pneumoniae . Mol Microbiol 51:917–924 [View Article][PubMed]
    [Google Scholar]
  25. Lama A., Kucknoor A., Mundodi V., Alderete J. F. ( 2009). Glyceraldehyde-3-phosphate dehydrogenase is a surface-associated, fibronectin-binding protein of Trichomonas vaginalis . Infect Immun 77:2703–2711 [View Article][PubMed]
    [Google Scholar]
  26. Lind K., Benzon M. W., Jensen J. S., Clyde W. A. Jr ( 1997). A seroepidemiological study of Mycoplasma pneumoniae infections in Denmark over the 50-year period 1946–1995. Eur J Epidemiol 13:581–586 [View Article][PubMed]
    [Google Scholar]
  27. Madureira P., Baptista M., Vieira M., Magalhães V., Camelo A., Oliveira L., Ribeiro A., Tavares D., Trieu-Cuot P. et al. ( 2007). Streptococcus agalactiae GAPDH is a virulence-associated immunomodulatory protein. J Immunol 178:1379–1387[PubMed] [CrossRef]
    [Google Scholar]
  28. May M., Papazisi L., Gorton T. S., Geary S. J. ( 2006). Identification of fibronectin-binding proteins in Mycoplasma gallisepticum strain R. Infect Immun 74:1777–1785 [View Article][PubMed]
    [Google Scholar]
  29. Miyata M. ( 2010). Unique centipede mechanism of Mycoplasma gliding. Annu Rev Microbiol 64:519–537 [View Article][PubMed]
    [Google Scholar]
  30. Nagata H., Iwasaki M., Maeda K., Kuboniwa M., Hashino E., Toe M., Minamino N., Kuwahara H., Shizukuishi S. ( 2009). Identification of the binding domain of Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase for Porphyromonas gingivalis major fimbriae. Infect Immun 77:5130–5138 [View Article][PubMed]
    [Google Scholar]
  31. Narita M. ( 2010). Pathogenesis of extrapulmonary manifestations of Mycoplasma pneumoniae infection with special reference to pneumonia. J Infect Chemother 16:162–169 [View Article][PubMed]
    [Google Scholar]
  32. Pancholi V., Chhatwal G. S. ( 2003). Housekeeping enzymes as virulence factors for pathogens. Int J Med Microbiol 293:391–401 [View Article][PubMed]
    [Google Scholar]
  33. Pancholi V., Fischetti V. A. ( 1992). A major surface protein on group A streptococci is a glyceraldehyde-3-phosphate-dehydrogenase with multiple binding activity. J Exp Med 176:415–426 [View Article][PubMed]
    [Google Scholar]
  34. Proft T., Herrmann R. ( 1994). Identification and characterization of hitherto unknown Mycoplasma pneumoniae proteins. Mol Microbiol 13:337–348 [View Article][PubMed]
    [Google Scholar]
  35. Purves J., Cockayne A., Moody P. C. E., Morrissey J. A. ( 2010). Comparison of the regulation, metabolic functions, and roles in virulence of the glyceraldehyde-3-phosphate dehydrogenase homologues GapA and GapB in Staphylococcus aureus . Infect Immun 78:5223–5232 [View Article][PubMed]
    [Google Scholar]
  36. Regula J. T., Boguth G., Görg A., Hegermann J., Mayer F., Frank R., Herrmann R. ( 2001). Defining the mycoplasma ‘cytoskeleton’: the protein composition of the Triton X-100 insoluble fraction of the bacterium Mycoplasma pneumoniae determined by 2-D gel electrophoresis and mass spectrometry. Microbiology 147:1045–1057[PubMed]
    [Google Scholar]
  37. Schmidl S. R., Gronau K., Pietack N., Hecker M., Becher D., Stülke J. ( 2010). The phosphoproteome of the minimal bacterium Mycoplasma pneumoniae: analysis of the complete known Ser/Thr kinome suggests the existence of novel kinases. Mol Cell Proteomics 9:1228–1242 [View Article][PubMed]
    [Google Scholar]
  38. Schurwanz N., Jacobs E., Dumke R. ( 2009). Strategy to create chimeric proteins derived from functional adhesin regions of Mycoplasma pneumoniae for vaccine development. Infect Immun 77:5007–5015 [View Article][PubMed]
    [Google Scholar]
  39. Søgaard I. Z., Boesen T., Mygind T., Melkova R., Birkelund S., Christiansen G., Schierup M. H. ( 2002). Recombination in Mycoplasma hominis . Infect Genet Evol 1:277–285 [View Article][PubMed]
    [Google Scholar]
  40. Tunio S. A., Oldfield N. J., Ala’Aldeen D. A., Wooldridge K. G., Turner D. P. ( 2010a). The role of glyceraldehyde 3-phosphate dehydrogenase (GapA-1) in Neisseria meningitidis adherence to human cells. BMC Microbiol 10:280 [View Article][PubMed]
    [Google Scholar]
  41. Tunio S. A., Oldfield N. J., Berry A., Ala’Aldeen D. A., Wooldridge K. G., Turner D. P. J. ( 2010b). The moonlighting protein fructose-1, 6-bisphosphate aldolase of Neisseria meningitidis: surface localization and role in host cell adhesion. Mol Microbiol 76:605–615 [View Article][PubMed]
    [Google Scholar]
  42. von Baum H., Welte T., Marre R., Suttorp N., Lück C., Ewig S. ( 2009). Mycoplasma pneumoniae pneumonia revisited within the German Competence Network for Community-acquired pneumonia (CAPNETZ). BMC Infect Dis 9:62 [View Article][PubMed]
    [Google Scholar]
  43. Waites K. B., Talkington D. F. ( 2004). Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev 17:697–728 [View Article][PubMed]
    [Google Scholar]
  44. Wilkins J. C., Beighton D., Homer K. A. ( 2003). Effect of acidic pH on expression of surface-associated proteins of Streptococcus oralis . Appl Environ Microbiol 69:5290–5296 [View Article][PubMed]
    [Google Scholar]
  45. Yavlovich A., Rechnitzer H., Rottem S. ( 2007). α-enolase resides on the cell surface of Mycoplasma fermentans and binds plasminogen. Infect Immun 75:5716–5719 [View Article][PubMed]
    [Google Scholar]
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