Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1975 Jun;122(3):1375–1386. doi: 10.1128/jb.122.3.1375-1386.1975

Cellular localization of lipoteichoic acid in Streptococcus faecalis.

R Joseph, G D Shockman
PMCID: PMC246194  PMID: 807556

Abstract

The release of lipoteichoic acid and mesosomal vesicles to the supernatant buffer during the formation of spherical, osmotically fragile bodies was studied using Streptococcus faecalis ATCC 9790. Autolytic N-acetylmuramidase action was permitted to take place in exponential-phase cells incubated in a buffer which provides an exceptional degree of osmotic stabilization. Both lipoteichoic acid and mesosomal vesicles were relatively rapidly released to the supernatant buffer. Most of the cellular content of lipoteichoic acid (and mesosomal vesicles) was found in the supernatant buffer at incubation times when the cells still retained over 75% of their cell wall. [14-C]- or [3-H]glycerol was used as a label for both cellular lipoteichoic acids and lipid-glycerol. Glycerol in lipoteichoic acid was quantitated after phenol-water and chloroform-methanol treatments and identified by products of acid hydrolysis and its ability to be precipitated by (i) antibodies specific for the polyglycerol-phosphate backbone, (ii) antibodies to the streptococcal group D antigen, and (iii) concanavalin A. Evidence was obtained that lipoteichoic acid was not associated with isolated mesosomal vesicles. Centrifugation of supernates at 200,000 X g sedimented membranous (mesosomal) vesicles and nearly all of the lipid-glycerol present, whereas essentially all of the lipoteichoic acid remained in the supernatant. The sedimented mesosomal vesicles differed from protoplast membrane in their higher lipid-phosphorus to protein ratio and in the absence of detectable levels of two enzymatic activities found in protoplast membranes, adenosine triphosphatase and polynucleotide phosphorylase. Both types of membranes were found to contain DD-carboxypeptidase and LD-transpeptidase activities at nearly the same specific activities. No evidence was obtained for the association of autolytic N-acetylmuramidase activity with either type of membrane preparation.

Full text

PDF
1375

Images in this article

1381Image
on p.1381

Selected References

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

  1. ABRAMS A., McNAMARA P., JOHNSON F. B. Adenosine triphosphatase in isolated bacterial cell membranes. J Biol Chem. 1960 Dec;235:3649–3662. [PubMed] [Google Scholar]
  2. Baddiley J. Teichoic acids in cell walls and membranes of bacteria. Essays Biochem. 1972;8:35–77. [PubMed] [Google Scholar]
  3. Burger M. M. Teichoic acids: antigenic determinants, chain separation, and their location in the cell wall. Proc Natl Acad Sci U S A. 1966 Sep;56(3):910–917. doi: 10.1073/pnas.56.3.910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Coyette J., Perkins H. R., Polacheck I., Shockman G. D., Ghuysen J. M. Membrane-bound DD-carboxypeptidase and LD-transpeptidase of Streptococcus faecalis ATCC 9790. Eur J Biochem. 1974 May 15;44(2):459–468. doi: 10.1111/j.1432-1033.1974.tb03504.x. [DOI] [PubMed] [Google Scholar]
  5. DISCHE Z., SHETTLES L. B. A specific color reaction of methylpentoses and a spectrophotometric micromethod for their determination. J Biol Chem. 1948 Sep;175(2):595–603. [PubMed] [Google Scholar]
  6. Ellar D. J., Thomas T. D., Posgate J. A. Properties of mesosomal membranes isolated from Micrococcus lysodeikticus and Bacillus megaterium. Biochem J. 1971 May;122(5):44P–45P. doi: 10.1042/bj1220044p. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ghuysen J. M., Bricas E., Leyh-Bouille M., Lache M., Shockman G. D. The peptide N alpha-(L-alanyl-D-isoglutaminyl)-N epsilon-(D-isoasparaginyl)-L-lysyl-D-alanine and the disaccharide N-acetylglucosaminyl-beta-1,4-N-acetylmuramic acid in cell wall peptidoglycan of Streptococcus faecalis strain ATCC 9790. Biochemistry. 1967 Aug;6(8):2607–2619. doi: 10.1021/bi00860a044. [DOI] [PubMed] [Google Scholar]
  8. HANES C. S., ISHERWOOD F. A. Separation of the phosphoric esters on the filter paper chromatogram. Nature. 1949 Dec 31;164(4183):1107-12, illust. doi: 10.1038/1641107a0. [DOI] [PubMed] [Google Scholar]
  9. HAY J. B., WICKEN A. J., BADDILEY J. The location of intracellular teichoic acids. Biochim Biophys Acta. 1963 Apr 2;71:188–190. doi: 10.1016/0006-3002(63)90999-5. [DOI] [PubMed] [Google Scholar]
  10. Heymann H., Manniello J. M., Barkulis S. S. Structure of streptococcal cell walls. V. Phosphate esters in the walls of group A Streptococcus pyogenes. Biochem Biophys Res Commun. 1967 Feb 21;26(4):486–491. doi: 10.1016/0006-291x(67)90574-8. [DOI] [PubMed] [Google Scholar]
  11. Higgins M. L., Pooley H. M., Shockman G. D. Site of initiation of cellular autolysis in Streptococcus faecalis as seen by electron microscopy. J Bacteriol. 1970 Aug;103(2):504–512. doi: 10.1128/jb.103.2.504-512.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Huff E., Cole R. M., Theodore T. S. Lipoteichoic acid localization in mesosomal vesicles of Staphylococcus aureus. J Bacteriol. 1974 Oct;120(1):273–281. doi: 10.1128/jb.120.1.273-281.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Joseph R., Shockman G. D. Autolytic formation of protoplasts (autoplasts) of Streptococcus faecalis 9790: release of cell wall, autolysin, and formation of stable autoplasts. J Bacteriol. 1974 May;118(2):735–746. doi: 10.1128/jb.118.2.735-746.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Knox K. W., Wicken A. J. Immunological properties of teichoic acids. Bacteriol Rev. 1973 Jun;37(2):215–257. doi: 10.1128/br.37.2.215-257.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROBERTS N. R., LEINER K. Y., WU M. L., FARR A. L. The quantitative histochemistry of brain. I. Chemical methods. J Biol Chem. 1954 Mar;207(1):1–17. [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Muñoz E., Nachbar M. S., Schor M. T., Salton M. R. Adenosinetriphosphatase of Micrococcus lysodeikticus: selective release and relationship to membrane structure. Biochem Biophys Res Commun. 1968 Aug 13;32(3):539–546. doi: 10.1016/0006-291x(68)90696-7. [DOI] [PubMed] [Google Scholar]
  18. Oppenheim J. D., Salton M. R. Localization and distribution of Micrococcus lysodeikticus membrane ATPase determined by ferritin labeling. Biochim Biophys Acta. 1973 Mar 16;298(2):297–322. doi: 10.1016/0005-2736(73)90360-x. [DOI] [PubMed] [Google Scholar]
  19. Owen P., Freer J. H. Isolation and properties of mesosomal membrane fractions from Micrococcus lysodeikticus. Biochem J. 1972 Oct;129(4):907–917. doi: 10.1042/bj1290907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reusch V. M., Jr, Burger M. M. Distribution of marker enzymes between mesosomal and protoplast membranes. J Biol Chem. 1974 Aug 25;249(16):5337–5345. [PubMed] [Google Scholar]
  21. Reusch V. M., Jr, Burger M. M. The bacterial mesosome. Biochim Biophys Acta. 1973 Apr 3;300(1):79–104. doi: 10.1016/0304-4157(73)90012-9. [DOI] [PubMed] [Google Scholar]
  22. Roth G. S., Shockman G. D., Daneo-Moore L. Balanced macromolecular biosynthesis in "protoplasts" of Streptococcus faecalis. J Bacteriol. 1971 Mar;105(3):710–717. doi: 10.1128/jb.105.3.710-717.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. SHOCKMAN G. D., SLADE H. D. THE CELLULAR LOCATION OF THE STREPTOCOCCAL GROUP D ANTIGEN. J Gen Microbiol. 1964 Dec;37:297–305. doi: 10.1099/00221287-37-3-297. [DOI] [PubMed] [Google Scholar]
  24. SMITH D. G., SHATTOCK P. M. THE CELLULAR LOCATION OF ANTIGENS IN STREPTOCOCCI OF GROUPS D, N AND Q. J Gen Microbiol. 1964 Jan;34:165–175. doi: 10.1099/00221287-34-1-165. [DOI] [PubMed] [Google Scholar]
  25. Salton M. R. Bacterial membranes. CRC Crit Rev Microbiol. 1971 May;1(1):161–197. doi: 10.3109/10408417109104480. [DOI] [PubMed] [Google Scholar]
  26. Sharpe M. E., Brock J. H., Knox K. W., Wicken A. J. Glycerol teichoic acid as a common antigenic factor in lactobacilli and some other gram-positive organisms. J Gen Microbiol. 1973 Jan;74(1):119–126. doi: 10.1099/00221287-74-1-119. [DOI] [PubMed] [Google Scholar]
  27. TOENNIES G., SHOCKMAN G. D., KOLB J. J. Differential effects of amino acid deficiencies on bacterial cytochemistry. Biochemistry. 1963 Mar-Apr;2:294–296. doi: 10.1021/bi00902a017. [DOI] [PubMed] [Google Scholar]
  28. TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]
  29. Theodore T. S., Popkin T. J., Cole R. M. The separation and isolation of plasma membranes and mesosomal vesicles from Staphylococcus aureus. Prep Biochem. 1971;1(3):233–248. doi: 10.1080/00327487108081942. [DOI] [PubMed] [Google Scholar]
  30. Toennies G., Das D. N., Feng F. New observations on the determination of bacterial lipid phosphorus. Can J Microbiol. 1968 Apr;14(4):484–485. doi: 10.1139/m68-079. [DOI] [PubMed] [Google Scholar]
  31. Toennies G., Feng F., Kolb J. J., Luttner P. M. Bacterial nucleate and phosphorus partition. Anal Biochem. 1965 Jun;11(3):473–496. doi: 10.1016/0003-2697(65)90067-9. [DOI] [PubMed] [Google Scholar]
  32. Toon P., Brown P. E., Baddiley J. The lipid-teichoic acid complex in the cytoplasmic membrane of Streptococcus faecalis N.C.I.B. 8191. Biochem J. 1972 Apr;127(2):399–409. doi: 10.1042/bj1270399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Van Driel D., Wicken A. J., Dickson M. R., Knox K. W. Cellular location of the lipoteichoic acids of Lactobacillus fermenti NCTC 6991 and Lactobacillus casei NCTC 6375. J Ultrastruct Res. 1973 Jun;43(5):483–497. doi: 10.1016/s0022-5320(73)90025-7. [DOI] [PubMed] [Google Scholar]
  34. WADE H. E., MORGAN D. M. Detection of phosphate esters on paper chromatograms. Nature. 1953 Mar 21;171(4351):529–530. doi: 10.1038/171529a0. [DOI] [PubMed] [Google Scholar]
  35. WICKEN A. J., BADDILEY J. Structure of intracellular teichoic acids from group D streptococci. Biochem J. 1963 Apr;87:54–62. doi: 10.1042/bj0870054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wicken A. J., Knox K. W. A serological comparison of the membrane teichoic acids from lactobacilli of different serological groups. J Gen Microbiol. 1971 Aug;67(2):251–254. doi: 10.1099/00221287-67-2-251. [DOI] [PubMed] [Google Scholar]
  37. Wicken A. J., Knox K. W. Lipoteichoic acids: a new class of bacterial antigen. Science. 1975 Mar 28;187(4182):1161–1167. doi: 10.1126/science.46620. [DOI] [PubMed] [Google Scholar]
  38. Wicken A. J., Knox K. W. Studies on the group F antigen of lactobacilli: isolation of a teichoic acid-lipid complex from Lactobacillus fermenti NCTC 6991. J Gen Microbiol. 1970 Mar;60(3):293–301. doi: 10.1099/00221287-60-3-293. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES