1887

Abstract

, an important aetiological agent of dental caries, is known to transport glucose via the phosphoenolpyruvate (PEP) phosphotransferase system (PTS). An alternative non-PTS glucose transport system in Ingbritt was suggested by the increased ATP-dependent phosphorylation of glucose and the presence of higher cellular concentration of free glucose in cells grown in continuous culture under PTS-repressed conditions compared to those resulting in optimal PTS activity. A method was developed for the preparation of membrane vesicles in order to study this system in the absence of PTS activity. These vesicles had very low activity of the cytoplasmic enzymes, glucokinase, pyruvate kinase and lactate dehydrogenase. This, coupled with the lack of glycolytic activity and the inability to transport glucose, suggested that the vesicles would also be deficient in PTS activity because of the absence of the general soluble PTS proteins, Enzyme I and HPr, required for the transport of all PTS sugars. Freeze fracture electron microscopy and membrane H-ATPase analysis indicated that over 90% of the vesicles had a right-side-out orientation. Vesicles from cells grown in continuous culture under PTS-dominant and PTS-repressed condition both exhibited glucose counterflow. This indicates the presence of a constitutive non-PTS carrier in the organism capable of transporting glucose and utilizing ATP for glucose phosphorylation. Analysis of growth yields of cells grown under PTS-repressed and PTS-optimal conditions suggests that ATP or an equivalent high energy molecule, must be involved in the actual transport process. This analysis is consistent with an ATP-binding protein model such as the transport system reported by R. R. B. Russell and coworkers (267, 4631-4637), but it does not exclude the possibilit of a separate permease for glucose.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-140-10-2639
1994-10-01
2021-05-12
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/10/mic-140-10-2639.html?itemId=/content/journal/micro/10.1099/00221287-140-10-2639&mimeType=html&fmt=ahah

References

  1. Bender G.R., Sutton S.V.W., Marquis R.E. Acid tolerance, proton permeabilities and membrane ATPases of oral streptococci. Infect Immtrn 1986; 53:331–338
    [Google Scholar]
  2. Bergmeyer H.U. Methods of Enzyme Analysis 1974 Deerfield Beach, Florida: Verlag Chemie International;
    [Google Scholar]
  3. Brown A.T., Wittenberger C.L. Fructose-1,6- diphosphate-dependent lactate dehydrogenase from a cariogenic streptococcus: purification and regulatory properties. J Bacterial 1972; 110:604–615
    [Google Scholar]
  4. Carlsson J. Metabolic activities of oral bacteria. In Textbook of Cariology 1986 Edited by Thylstrup A., Fejerskov O. Copenhagen : Munksgaard; pp 74–106
    [Google Scholar]
  5. Carlsson J., Griffith C.J. Fermentation products and bacterial yields in glucose-limited and nitrogen-limited cultures of streptococci. Arch Oral Biol 1974; l9:1105–1109
    [Google Scholar]
  6. Chapman R.L., Staehelin L.A. Freeze-fracture (-etch) electron microscopy. In Ultrastructure Techniques for Microorganisms 1986 Edited by Aldrich H.C., Todd W.J. New York: Plenum Press; pp 213–240
    [Google Scholar]
  7. Chassey B., Giuffrida A. Method for the lysis of Grampositive, asporogenous bacteria with lysozyme. Appl Environ Microbiol 1980; 39:153–158
    [Google Scholar]
  8. Dashper S.G., Reynolds E.C. Characterization of transmembrane movement of glucose and glucose analogues in Streptococcus S. mutans Ingbritt. J Bacteriol 1990; 172:556–563
    [Google Scholar]
  9. Dashper S.G., Reynolds E.C. Branched-chain amino acid transport in Streptococcus S. mutans Ingbritt. Oral Microbiol Immunol 1993; 8:167–171
    [Google Scholar]
  10. Eisenberg R.J., Lillmars K. A method for the gentle lysis of Streptococcus sanguis and Streptococcus S mutans. Biochem Biophys Res Commun 1975; 65:378–384
    [Google Scholar]
  11. Ellwood D.C., Hamilton I.R. Properties of Streptococcus S. mutans Ingbritt growing in limiting sucrose in a chemostat: repression of the phosphoenolpyruvate phosphotransferase transport system. Infect Immun 1982; 36:576–581
    [Google Scholar]
  12. Ellwood D., C, Phipps P.J., Hamilton I.R. Effect of growth rate and glucose concentration on the activity of the phosphoenolpyruvate phosphotransferase system in Streptococcus S mutans Ingbritt grown in continuous culture. Infect Immun 1979; 23:224–231
    [Google Scholar]
  13. Futai M., Kanazawa H. Structure and function of the proton-translocating adenosine triphosphatase (FoFj): biochemical and molecular biological approaches. Microbiol Rev 1983; 47:285–312
    [Google Scholar]
  14. Goodman H., Pollack J.J., Lacono V.J., Wong W., Shockman G. Peptidoglycan loss during hen egg white lysozyme- inorganic salt lysis of Streptococcus S mutans. J Bacteriol 1981; 146:755–763
    [Google Scholar]
  15. Gottschalk G. Bacterial Metabolism Springer-Verlag; 1978
    [Google Scholar]
  16. Hamada S., Slade H.D. Biology, immunology and cariogenicity of Streptococcus S mutans. Microbiol Rev 1980; 44:331–284
    [Google Scholar]
  17. Hamilton I.R. Growth of the oral ‘pathogen’, Streptococcus S. mutans in continuous culture reveals two glucose transport systems. In Continuous Culture 8: Biotechnology 1984 Edited by Dean A.C.R., Ellwood D.C., Evans C.G.T. Chichester : Ellis Horwood; Medicine and the Environment, pp 58–71
    [Google Scholar]
  18. Hamilton I.R. Effect of changing environment on sugar transport and metabolism by oral bacteria. In Sugar Transport and Metabolism by Gram-Positive Bacteria 1987 Edited by Reizer J., Peterkofsky A. Chichester: Ellis Horwood; pp 94–133
    [Google Scholar]
  19. Hamilton I.R. Maintenance of protonmotive force by Streptococcus S mutans and Streptococcus sobrinus during growth in continuous culture. Oral Microbiol Immunol 1990; 5:280–287
    [Google Scholar]
  20. Hamilton I.R., Buckley N.D. Adaptation of Streptococcus S mutans to acid tolerance. Oral Microbiol Immunol 1991; 6:65–71
    [Google Scholar]
  21. Hamilton I.R., Ellwood D.C. Effects of fluoride on carbohydrate metabolism by washed cells of Streptococcus S. mutans grown at various pH values in a chemostat. Infect Immun 1978; 19:434–442
    [Google Scholar]
  22. Hamilton I.R., St Martin E.J. Evidence for the involvement of protonmotive force in the transport of glucose by a mutant of Streptococcus S mutans strain DR0001 defective in the glucose phosphoenolpyruvate phosphotransferase system. Infect Immun 1982; 36:567–575
    [Google Scholar]
  23. Hamilton I.R., Phipps P.J., Ellwood D.C. Effect of growth rate and glucose concentration on the biochemical properties of Streptococcus S mutans Ingbritt in continuous culture. Infect Immun 1979; 26:861–869
    [Google Scholar]
  24. Hamilton I.R., Gauthier L., Desjardins B., Vadeboncoeur C. Concentration-dependent repression of the soluble and membrane components of the phosphoenolpyruvate: sugar phosphotransferase system of Streptococcus S. mutans by glucose. J Bacteriol 1989; 171:2942–2948
    [Google Scholar]
  25. Van Houte J. Bacterial specificity in the etiology of caries. Int Dent J 1986; 30:305–326
    [Google Scholar]
  26. Kaback H.R. Uptake of amino acids by ‘ ghosts ’ of mutant strains of E coli. Fed Proc 1960; 19:130
    [Google Scholar]
  27. Kaback H.R. The role of the phosphoenolpyruvate- phosphotransferase system in the transport of sugars by isolated membrane preparations of Escherichia coli. J Biol Chem 1968; 243:3711–3724
    [Google Scholar]
  28. Kaback H.R. Transport across isolated bacterial cytoplasmic membranes. Biochim Biophys Acta 1972; 265:367–416
    [Google Scholar]
  29. Kaback H.R. Transport studies in bacterial membrane vesicles. Science 1974; 186:882–892
    [Google Scholar]
  30. Khandelwal R.J., Hamilton I.R. Purification and properties of adenyl cyclase from Streptococcus salivarius. J Biol Chem 1971; 246:3297–3304
    [Google Scholar]
  31. Kingsley G.R., Getchell G. Direct ultra micro glucose oxidase method for the determination of glucose in biological fluids. Clin Chem 1960; 6:466–475
    [Google Scholar]
  32. Kobayashi H., Van Brunt J., Harold F.M. A TP-linked calcium transport in cells and membrane vesicles of Streptococcus faecalis. J Biol Chem 1978; 253:2085–2092
    [Google Scholar]
  33. Konings W.N., De Vrij W., Driessen A.J.M., Poolman B. Primary and secondary transport in Gram-positive bacteria. In Sugar Transport and Metabolism by Gram-Positive Bacteria 1987 Edited by Reizer J., Peterkofsky A. Chichester: Eillis Horwood; pp 270–294
    [Google Scholar]
  34. Maloney P.C., Kashket E.R., Wilson T.H. Methods for studying transport in bacteria. Methods Membr Biol 1975; 5:1–49
    [Google Scholar]
  35. Martin N.L., Beveridge T.J. Gentamicin interaction with the Pseudomonas aeruginosa cell envelope. Antimicrob Agents Chemother 1986; 29:1079–1087
    [Google Scholar]
  36. Meadow N.D., Fox D.K., Roseman S. The bacterial phosphoenolpyruvate: glycose phosphotransferase system. Annu Rev Biochem 1990; 59:497–542
    [Google Scholar]
  37. Otto R., Klont B., Ten Brink B., Konings W.N. The phosphate potential, adenylate charge and protonmotive force in growing cells of Streptococcus cremoris. Arch Microbiol 1984; 139:338–343
    [Google Scholar]
  38. Peterson G.L. Determination of total protein. Methods Enzymol 1983; 91:95–119
    [Google Scholar]
  39. Porter V.E., Chassey B.M., Holmlund C.E. Partial purification and properties of a specific glucokinase from Streptococcus S. mutans SL-1. Biochim Biophys Acta 1980; 611:289–298
    [Google Scholar]
  40. Postma P.W., Stock J.B. Enzymes II of the phosphotransferase system do not catalyze sugar transport in the absence of phosphorylation. J Bacteriol 1980; 141:476–484
    [Google Scholar]
  41. Postma P.W., Lengeler J.W., Jacobson G.R. Phosphoenolpyruvate: carbohydrate phosphotransferase system of bacteria. Microbiol Rev 1993; 57:543–594
    [Google Scholar]
  42. Robillard G.T. Functional reconstitution of the purified phosphoenolpyruvate-dependent mannitol specific transport system of Escherichia coli in phospholipid vesicles: coupling between transport and phosphorylation. J Bacteriol 1987; 172:7119–7125
    [Google Scholar]
  43. Roseman S. The transport of carbohydrates by a bacterial phosphotransferase system. J Gen Physiol 1969; 54:138–184
    [Google Scholar]
  44. Ruijter G.C.G., Van Meurs G., Verwey M.A., Postma P.W., Van Dam K. Analysis of mutations that uncouple transport from phosphorylation in Enzyme IIGlcof the Escherichia coli phosphoenolpyruvate phosphotransferase system. J Bacteriol 1992; 174:2843–2850
    [Google Scholar]
  45. Russell J.B. Low-aflflnity, high-capacity system of glucose transport in the ruminal bacterium Streptococcus bovis: evidence for a mechanism of facilitated diffusion. Appl Environ Microbiol 1990; 56:3304–3307
    [Google Scholar]
  46. Russell R.R.B., Aduse-Opoku J., Sutcliffe I.C., Tao L., Ferretti J.J. A binding protein-dependent transport system in Streptococcus S. mutans responsible for multiple sugar metabolism. J Biol Chem 1992; 267:4631–4637
    [Google Scholar]
  47. Saier M. Jr Mechanisms and Regulation of Carbohydrate Transport in Bacteria 1985 New York: Academic Press, Inc;
    [Google Scholar]
  48. Scholler M., Klein J.P., Sommer P., Frank R. Protoplast and cytoplasmic membrane preparations from Streptococcus sanguis and Streptococcus S. mutans. J Gen Microbiol 1983; 129:3271–3279
    [Google Scholar]
  49. Siegel J.L., Hurst S.F., Liberman E.S., Coleman S.E., Bleiweis A.S. Mutanolysin-induced spheroplasts of Streptococcus S. mutans are true protoplasts. Infect Immun 1981; 31:808–815
    [Google Scholar]
  50. Stempek J.G., Ward R.T. An improved method for electron microscopy. J Cell Biol 1964; 22:697–701
    [Google Scholar]
  51. Tao L., Sutcliffe I.C., Russell R.R.B., Ferretti J.J. Transport of sugars, including sucrose, by the msm transport system of Streptococcus S. mutans. J Dent Rir 1993; 267:4631–4637
    [Google Scholar]
  52. Vadeboncoeur C. Structure and properties of the phosphoenolpyruvate: glucose phosphotransferase system of oral streptococci. Can J Microbiol 1984; 30:495–502
    [Google Scholar]
  53. Vadeboncoeur C., Thibault L., Neron S., Halvorson H., Hamilton I.R. Effect of growth conditions on levels of components of the phosphoenolpyruvate: sugar phosphotransferase system in Streptococcus S. mutans and Streptococcus sobrinus grown in continuous culture. J Bacteriol 1987; 169:5686–5691
    [Google Scholar]
  54. Vadeboncoeur C., St Martin S., Brochu D., Hamilton I.R. Effect of growth rate and pH on intracellular levels and activities of the components of the phosphoenolpyruvate: sugar phosphotransferase system in Streptococcus S mutans Ingbritt. Infect Immun 1991; 59:900–906
    [Google Scholar]
  55. Venable J.H., Coggeshell R. A simplified lead citrate stain for use in electron microscopy. J Cell Biol 1965; 25:407–412
    [Google Scholar]
  56. Yamada T., Carlsson J. Glucose-6-phosphate dependent pyruvate kinase in Streptococcus S. mutans. J Bacteriol 1975; 124:562–563
    [Google Scholar]
  57. Yuan L.C., Gulyas B.J. An improved method for processing single cells for E.M. utilizing agarose. Anat Rec 1981; 201:273–281
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-140-10-2639
Loading
/content/journal/micro/10.1099/00221287-140-10-2639
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error