1887

Abstract

HPr is a low-molecular-mass phosphocarrier protein of the bacterial phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS) found in the cytoplasm or associated with the inner surface of the cytoplasmic membrane. Treatment of cells with a Sorvall Omnimixer, a technique used to extract cell surface components, resulted in the extraction of a major protein with a molecular mass of 9 kDa. Several lines of evidence suggested that this protein was HPr: (i) the protein showed homology over the first 35 N-terminal amino acid residues with the HPrs of and , including the signature sequence for the site of PEP-dependent phosphorylation; (ii) it cross-reacted with the anti-HPr antibody preparation; (iii) it could be phosphorylated by enzyme 1 at the expense of PEP, and by a membrane-associated kinase at the expense of ATP; and (iv) it possessed phosphocarrier activity when used as a source of HPr in an PTS assay. The data suggested that a portion of the cellular HPr is associated with the external cell surface in , a result that was confirmed by immunogold electron microscopy. The cellular HPr of consisted of two forms that could be distinguished by the presence or the absence of the N-terminal methionine. Amino acid sequence analysis indicated that the cell-surface-associated HPr of lacked the N-terminal methionine residue.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-142-4-837
1996-04-01
2021-10-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/4/mic-142-4-837.html?itemId=/content/journal/micro/10.1099/00221287-142-4-837&mimeType=html&fmt=ahah

References

  1. Arends J.P., Zanen H.C. Meningitis caused by Streptococcus suis in humans. Rep Infect Dis 1988; 10:131–137
    [Google Scholar]
  2. Bentley R.W., Leigh J.A., Collins M.D. Intragenic structure of Streptococcus based on comparative analysis of small-subunit rRNA sequences. Int J Sjst Bacteriol 1991; 41:487–494
    [Google Scholar]
  3. Boyd D.A., Cvitkovitch D.G., Hamilton I.R. Sequence and expression of the genes for HPr (ptsH) and enzyme I (ptsl) of the phosphoenolpyruvate-dependent phosphotransferase transport system from Streptococcus mutans. Infect Immun 1994; 62:1156–1165
    [Google Scholar]
  4. Deutscher J., Sossna G., Gonzy-Treboul G. Regulatory functions of the phosphocarrier protein HPr of the phosphoenolpyruvate-dependent phosphotransferase system in Grampositive bacteria. FEMS Microbiol Eett 1989; 63:67–174
    [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids R« 1984; 12:387–395
    [Google Scholar]
  6. Dills S.S., Apperson A., Schmidt M., Saier M.H. Jr Carbohydrate transport in bacteria. Microbiol Rep 1980; 44:385–418
    [Google Scholar]
  7. Gagnon G., Vadeboncoeur C., Frenette M. Phosphotransferase system of Streptococcus salivarius: characterization of the ptsH gene and its product. Gene 1993; 136:27–34
    [Google Scholar]
  8. Gauthier L., Mayrand D., Vadeboncoeur C. Isolation of a novel protein involved in the transport of fructose by an inducible phosphoenolpy ruvate fructose phosphotransferase system in Streptococcus mutans. J Bacteriol 1984; 160:755–763
    [Google Scholar]
  9. Gerlach D., Alouf H., Moraved L., Pavlik M., Kohler W. The characterization of two new low molecular weight proteins (LMPs) from Streptococcus pyogenes. Zentralbl Bakteriol Mikrobiol Hyg 1992; 277:1–9
    [Google Scholar]
  10. 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]
  11. Hamilton I.R., Gauthier L., Desjardins B., Vadeboncoeur C. Concentration-dependent repression of the soluble and membrane components of the Streptococcus mutans phosphoenol-pyruvate: sugar phosphotransferase system by glucose. J Bacteriol 1989; 171:2942–2948
    [Google Scholar]
  12. Hawkes R. Identification of concavalin A-binding proteins after sodium dodecyl sulfate-gel electrophoresis and protein blotting. Anal Biochem 1982; 123:143–146
    [Google Scholar]
  13. Heckels J.E., Virji M. Separation and purification of surface components. In Bacterial Cell Surface Techniques 1988 Edited by Hancock I.C., Poxton I.R. Chichester: John Wiley; pp 67–135
    [Google Scholar]
  14. Hewick R.M., Humkapiller M.W., Hood L.E., Dreyer W.J. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem 1980; 256:7990–7997
    [Google Scholar]
  15. Higgins R., Gottschalk M., Mittal K.R., Beaudoin M. Streptococcus suis infections in swine A 16 month study. Can J Vet R« 1990; 54:170–173
    [Google Scholar]
  16. Hommez J., Devriese L.A., Henrichsen J., Castryck F. Identification and characterization of Streptococcus suis. Vet Microbiol 1986; 11:349–355
    [Google Scholar]
  17. Jenkinson H.F. Properties of a phosphocarrier protein (HPr) extracted from intact cells of Streptococcus sanguis. J Gen Microbiol 1989; 135:3183–3197
    [Google Scholar]
  18. Martensen T.M. Chemical properties, isolation and analysis of O-phosphates in proteins. Methods Enzymol 1984; 107:3–23
    [Google Scholar]
  19. Meadow N.D., f Fox D.K., Roseman S. The bacterial phosphoenolpyruvate:glycose phosphotransferase system. Annu Rev Biochem 1990; 5:497–542
    [Google Scholar]
  20. Pelletier M., Frenette M., Vadeboncoeur C. Distribution of proteins similar to III“an and III)1an of the Streptococcus salivarius phosphoenolpyruvate: mannose-glucose phosphotransferase system among oral and nonoral bacteria. J Bacteriol 1995; 177:2270–2275
    [Google Scholar]
  21. Postma P.W., Lengeler J.W., Jacobson G. Phosphoenolpyruvate: carbohydrate phosphotransferase systems of bacteria. Microbiol Rev 1993; 57:543–594
    [Google Scholar]
  22. Reizer J., Deutscher J., Saier M.H. Jr Metabolitesensitive, ATP-dependent, protein kinase-catalysed phosphorylation of HPr, a phosphocarrier protein of the phosphotransferase system in Gram-positive bacteria. Biochimie 1989; 71:989–996
    [Google Scholar]
  23. Reizer J., Hoischen C., Reizer A., Pham T.N., Saier M.H. Jr Sequence analyses and evolutionary relationships among the energy-coupling proteins Enzyme I and HPr of the bacterial phosphoenolpyruvate: sugar phosphotranferase system. Protein Sci 1993a; 2:506–521
    [Google Scholar]
  24. Reizer J., Romano A.H., Deutscher J. The role of phosphorylation of HPr, a phosphocarrier protein of the phosphotransferase system, in the regulation of carbon metabolism in Gram-positive bacteria. J Cell Biochem 1993b; 51:19–24
    [Google Scholar]
  25. Robitaille D., Gauthier L., Vadeboncoeur C. The presence of two forms of the phosphocarrier protein HPr of the phosphoenolpyruvate:sugar phosphotransferase system in streptococci. Biochimie 1991; 73:573–581
    [Google Scholar]
  26. Russell R.R.B. Isolation and purification of proteins linked to the cell wall in Gram-positive bacteria. In Bacterial Cell Surface Techniques 1988 Edited by Hancock I.C., Poxton I.R. Chichester: John Wiley; pp 104–110
    [Google Scholar]
  27. Saier M.H. Jr, Reizer J. Proposed uniform nomenclature for the proteins and protein domains of the bacterial phosphoenolpyruvate: sugar phosphotransferase system. J Bacteriol 1992; 174:1433–1438
    [Google Scholar]
  28. Saier M.H. Jr, Cox D.F., Feucht B.U., Novotny M.J. Evidence for the functional association of Enzyme I and HPr of the phosphoenolpyruvate-sugar phosphotranferase system with the membrane in sealed vesicles of Escherichia coli. J Cell Biochem 1982; 18:231–238
    [Google Scholar]
  29. Simonen M., Palva I. Protein secretion in Bacillus species. Microbiol Rev 1993; 57:109–137
    [Google Scholar]
  30. Sutcliffe I.G., Hogg S.D., Russell R.R.B. Identification of Streptococcus mutans antigen D as the HPr protein of the sugar-phosphotransferase transport system. FEMS Microbiol Lett 1993; 107:67–70
    [Google Scholar]
  31. Swank R.T., Munkres K.D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecylsulfate. Anal Biochem 1971; 39:462–477
    [Google Scholar]
  32. Thompson J. Sugar transport in the lactic acid bacteria. In Sugar Transport and Metabolism bj Gram-positive Bacteria 1987 Edited by Reizer J., Peterkofsky A. Chichester: Ellis Horwood; pp 13–38
    [Google Scholar]
  33. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76:4350–4354
    [Google Scholar]
  34. Trottier S., Higgins R., Brochu G., Gottschalk M. A case of human endocarditis due to Streptococcus suis in North America. Rev Infect Dis 1991; 13:1251–1252
    [Google Scholar]
  35. Vadeboncoeur C. HPr: Heteromorphous Protein. Res Microbiol 1995; 146:525–530
    [Google Scholar]
  36. Vadeboncoeur C., Brochu D., Reizer J. Quantitative determination of the intracellular concentration of the various forms of HPr, a phosphocarrier protein of the phosphoenolpyruvate: sugar phosphotransferase system in growing cells of oral streptococci. Anal Biochem 1991a; 196:24–30
    [Google Scholar]
  37. Vadeboncoeur C., Konishi Y., Dumas F., Gauthier L., Frenette M. HPr polymorphism in oral streptococci is caused by the partial removal of the N-terminal methionine. Biochimie 1991b; 73:1427–1430
    [Google Scholar]
  38. Vadeboncoeur G., Brochu D., Trahan L., Fradette J., Gingras S. Amino-terminal methionine processing of the protein HPr in Streptococcus salivarius grown in continuous culture. FEMS Microbiol Lett 1993; 111:197–202
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-142-4-837
Loading
/content/journal/micro/10.1099/00221287-142-4-837
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