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. 1991 Feb;173(3):1041–1050. doi: 10.1128/jb.173.3.1041-1050.1991

Localization of the Mycoplasma pneumoniae cytadherence-accessory proteins HMW1 and HMW4 in the cytoskeletonlike Triton shell.

M K Stevens 1, D C Krause 1
PMCID: PMC207223  PMID: 1899414

Abstract

The location of the cytadherence-accessory high-molecular weight proteins 1 and 4 (HMW1/4) within Mycoplasma pneumoniae cells has been studied by both biochemical and electron microscopic techniques. Peptide mapping studies demonstrated that HMW1/4 share almost identical peptide profiles, suggesting that the two proteins are structurally related. Examination of thin sections of M. pneumoniae with antibodies to HMW1/4 and colloidal gold particles revealed distinct labeling of the filamentous extensions of the mycoplasma cells. Labeling was absent on thin sections of a cytadherence-deficient variant lacking HMW1/4. HMW1/4 partitioned in the detergent-insoluble fraction following Triton X-100 extraction, and analysis by sucrose density gradient centrifugation suggested that HMW1/4 are part of a high-molecular-weight, multiprotein complex. These results were confirmed by immunogold labeling of Triton X-100-extracted M. pneumoniae cells incubated with antibodies to HMW1/4: gold particles bound in specific clusters to detergent-insoluble filaments. Finally, immunogold labeling of whole cells revealed that HMW1/4 are exposed on the cell surface, although to a lesser degree than on the cell interior. These findings indicate that HMW1/4 are membrane proteins associated with the cytoskeletonlike triton shell of M. pneumoniae and localized primarily in the filamentous extensions of the mycoplasma cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baseman J. B., Cole R. M., Krause D. C., Leith D. K. Molecular basis for cytadsorption of Mycoplasma pneumoniae. J Bacteriol. 1982 Sep;151(3):1514–1522. doi: 10.1128/jb.151.3.1514-1522.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baseman J. B., Morrison-Plummer J., Drouillard D., Puleo-Scheppke B., Tryon V. V., Holt S. C. Identification of a 32-kilodalton protein of Mycoplasma pneumoniae associated with hemadsorption. Isr J Med Sci. 1987 May;23(5):474–479. [PubMed] [Google Scholar]
  3. Bennett V., Stenbuck P. J. The membrane attachment protein for spectrin is associated with band 3 in human erythrocyte membranes. Nature. 1979 Aug 9;280(5722):468–473. doi: 10.1038/280468a0. [DOI] [PubMed] [Google Scholar]
  4. Bennett V. The membrane skeleton of human erythrocytes and its implications for more complex cells. Annu Rev Biochem. 1985;54:273–304. doi: 10.1146/annurev.bi.54.070185.001421. [DOI] [PubMed] [Google Scholar]
  5. Biberfeld G., Biberfeld P. Ultrastructural features of Mycoplasma pneumoniae. J Bacteriol. 1970 Jun;102(3):855–861. doi: 10.1128/jb.102.3.855-861.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bredt W., Feldner J., Kahane I. Adherence of mycoplasmas to cells and inert surfaces: phenomena, experimental models and possible mechanisms. Isr J Med Sci. 1981 Jul;17(7):586–588. [PubMed] [Google Scholar]
  7. Collier A. M., Clyde W. A., Jr Appearance of Mycoplasma pneumoniae in lungs of experimentally infected hamsters and sputum from patients with natural disease. Am Rev Respir Dis. 1974 Dec;110(6):765–773. doi: 10.1164/arrd.1974.110.6P1.765. [DOI] [PubMed] [Google Scholar]
  8. Collier A. M., Clyde W. A. Relationships Between Mycoplasma pneumoniae and Human Respiratory Epithelium. Infect Immun. 1971 May;3(5):694–701. doi: 10.1128/iai.3.5.694-701.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crossley R., Holberton D. V. Characterization of proteins from the cytoskeleton of Giardia lamblia. J Cell Sci. 1983 Jan;59:81–103. doi: 10.1242/jcs.59.1.81. [DOI] [PubMed] [Google Scholar]
  10. Feldner J., Göbel U., Bredt W. Mycoplasma pneumoniae adhesin localized to tip structure by monoclonal antibody. Nature. 1982 Aug 19;298(5876):765–767. doi: 10.1038/298765a0. [DOI] [PubMed] [Google Scholar]
  11. Göbel U., Speth V., Bredt W. Filamentous structures in adherent Mycoplasma pneumoniae cells treated with nonionic detergents. J Cell Biol. 1981 Nov;91(2 Pt 1):537–543. doi: 10.1083/jcb.91.2.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Haest C. W. Interactions between membrane skeleton proteins and the intrinsic domain of the erythrocyte membrane. Biochim Biophys Acta. 1982 Dec;694(4):331–352. doi: 10.1016/0304-4157(82)90001-6. [DOI] [PubMed] [Google Scholar]
  13. Hansen E. J., Wilson R. M., Baseman J. B. Isolation of mutants of Mycoplasma pneumoniae defective in hemadsorption. Infect Immun. 1979 Mar;23(3):903–906. doi: 10.1128/iai.23.3.903-906.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hansen E. J., Wilson R. M., Baseman J. B. Two-dimensional gel electrophoretic comparison of proteins from virulent and avirulent strains of Mycoplasma pneumoniae. Infect Immun. 1979 May;24(2):468–475. doi: 10.1128/iai.24.2.468-475.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hansen E. J., Wilson R. M., Clyde W. A., Jr, Baseman J. B. Characterization of hemadsorption-negative mutants of Mycoplasma pneumoniae. Infect Immun. 1981 Apr;32(1):127–136. doi: 10.1128/iai.32.1.127-136.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hayflick L. Tissue cultures and mycoplasmas. Tex Rep Biol Med. 1965 Jun;23(Suppl):285+–285+. [PubMed] [Google Scholar]
  17. Hu P. C., Cole R. M., Huang Y. S., Graham J. A., Gardner D. E., Collier A. M., Clyde W. A., Jr Mycoplasma pneumoniae infection: role of a surface protein in the attachment organelle. Science. 1982 Apr 16;216(4543):313–315. doi: 10.1126/science.6801766. [DOI] [PubMed] [Google Scholar]
  18. Hu P. C., Collier A. M., Baseman J. B. Surface parasitism by Mycoplasma pneumoniae of respiratory epithelium. J Exp Med. 1977 May 1;145(5):1328–1343. doi: 10.1084/jem.145.5.1328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kahane I., Tucker S., Leith D. K., Morrison-Plummer J., Baseman J. B. Detection of the major adhesin P1 in triton shells of virulent Mycoplasma pneumoniae. Infect Immun. 1985 Dec;50(3):944–946. doi: 10.1128/iai.50.3.944-946.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Krause D. C., Baseman J. B. Inhibition of mycoplasma pneumoniae hemadsorption and adherence to respiratory epithelium by antibodies to a membrane protein. Infect Immun. 1983 Mar;39(3):1180–1186. doi: 10.1128/iai.39.3.1180-1186.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Krause D. C., Baseman J. B. Mycoplasma pneumoniae proteins that selectively bind to host cells. Infect Immun. 1982 Jul;37(1):382–386. doi: 10.1128/iai.37.1.382-386.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Krause D. C., Leith D. K., Baseman J. B. Reacquisition of specific proteins confers virulence in Mycoplasma pneumoniae. Infect Immun. 1983 Feb;39(2):830–836. doi: 10.1128/iai.39.2.830-836.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Krause D. C., Leith D. K., Wilson R. M., Baseman J. B. Identification of Mycoplasma pneumoniae proteins associated with hemadsorption and virulence. Infect Immun. 1982 Mar;35(3):809–817. doi: 10.1128/iai.35.3.809-817.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Lipman R. P., Clyde W. A., Jr, Denny F. W. Characteristics of virulent, attenuated, and avirulent Mycoplasma pneumoniae strains. J Bacteriol. 1969 Nov;100(2):1037–1043. doi: 10.1128/jb.100.2.1037-1043.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lux S. E. Dissecting the red cell membrane skeleton. Nature. 1979 Oct 11;281(5731):426–429. doi: 10.1038/281426a0. [DOI] [PubMed] [Google Scholar]
  27. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  28. Meng K. E., Pfister R. M. Intracellular structures of Mycoplasma pneumoniae revealed after membrane removal. J Bacteriol. 1980 Oct;144(1):390–399. doi: 10.1128/jb.144.1.390-399.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Niggli V., Burger M. M. Interaction of the cytoskeleton with the plasma membrane. J Membr Biol. 1987;100(2):97–121. doi: 10.1007/BF02209144. [DOI] [PubMed] [Google Scholar]
  30. Oakley B. R., Kirsch D. R., Morris N. R. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem. 1980 Jul 1;105(2):361–363. doi: 10.1016/0003-2697(80)90470-4. [DOI] [PubMed] [Google Scholar]
  31. Powell D. A., Hu P. C., Wilson M., Collier A. M., Baseman J. B. Attachment of Mycoplasma pneumoniae to respiratory epithelium. Infect Immun. 1976 Mar;13(3):959–966. doi: 10.1128/iai.13.3.959-966.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Roberts D. D., Olson L. D., Barile M. F., Ginsburg V., Krivan H. C. Sialic acid-dependent adhesion of Mycoplasma pneumoniae to purified glycoproteins. J Biol Chem. 1989 Jun 5;264(16):9289–9293. [PubMed] [Google Scholar]
  33. Sobeslavsky O., Prescott B., Chanock R. M. Adsorption of Mycoplasma pneumoniae to neuraminic acid receptors of various cells and possible role in virulence. J Bacteriol. 1968 Sep;96(3):695–705. doi: 10.1128/jb.96.3.695-705.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tjian R., Stinchcomb D., Losick R. Antibody directed against Bacillus subtilis rho factor purified by sodium dodecyl sulfate slab gel electrophoresis. Effect on transcription by RNA polymerase in crude extracts of vegetative and sporulating cells. J Biol Chem. 1975 Nov 25;250(22):8824–8828. [PubMed] [Google Scholar]
  35. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]

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