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. 1985 Dec;164(3):1337–1349. doi: 10.1128/jb.164.3.1337-1349.1985

Use of resistant mutants to study the interaction of triton X-100 with Staphylococcus aureus.

D Raychaudhuri, A N Chatterjee
PMCID: PMC219335  PMID: 2866176

Abstract

Staphylococcus aureus mutants resistant to the nonionic detergent Triton X-100, isolated from the wild-type strain H and the autolysin-deficient strain RUS3, could grow and divide in broth containing 5% (vol/vol) Triton X-100, while growth of the parental strains was markedly inhibited above the critical micellar concentration (0.02%) of the detergent. Growth-inhibitory concentrations of Triton X-100 killed wild-type cells without demonstrable cellular lysis. Triton X-100 stimulated autolysin activity of S. aureus cells under nongrowing conditions, and this lytic response was markedly reduced in energy-poisoned cells. In contrast, the detergent had no effect on the activity of autolysins in cell-free systems, and growth in the presence of Triton X-100 did not alter either the cellular autolysin activity or the susceptibility of cell walls to exogenous lytic enzymes. Treatment with either Triton X-100 or penicillin G in the growth medium stimulated release of predominantly acylated intracellular lipoteichoic acid and sensitized staphylococci to Triton X-100-induced autolysis. There was no significant difference in the cell wall and membrane compositions or Triton X-100 binding between the parental strains and the resistant mutants. The resistant mutant TXR1, derived from S. aureus H, had a higher level of L-alpha-glycerophosphate dehydrogenase activity, and its oxygen uptake was more resistant to inhibition by a submicellar concentration (0.008%) of Triton X-100. Growth in the presence of subinhibitory concentrations of Triton X-100 rendered S. aureus H cells phenotypically resistant to the detergent and greatly stimulated the level of oxygen uptake. Membranes isolated from such cells exhibited enhanced activity of the respiratory enzymes succinic dehydrogenase and L-alpha-glycerophosphate dehydrogenase.

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Selected References

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  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. BROWDER H. P., ZYGMUNT W. A., YOUNG J. R., TAVORMINA P. A. LYSOSTAPHIN: ENZYMATIC MODE OF ACTION. Biochem Biophys Res Commun. 1965 Apr 23;19:383–389. doi: 10.1016/0006-291x(65)90473-0. [DOI] [PubMed] [Google Scholar]
  4. Chalvardjian A. M. Fatty acids of brown and yellow fat in rats. Biochem J. 1964 Mar;90(3):518–521. doi: 10.1042/bj0900518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chatterjee A. N. Use of bacteriophage-resistant mutants to study the nature of the bacteriophage receptor site of Staphylococcus aureus. J Bacteriol. 1969 May;98(2):519–527. doi: 10.1128/jb.98.2.519-527.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chatterjee A. N., Wong W., Young F. E., Gilpin R. W. Isolation and characterization of a mutant of Staphylococcus aureus deficient in autolytic activity. J Bacteriol. 1976 Mar;125(3):961–967. doi: 10.1128/jb.125.3.961-967.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cleveland R. F., Daneo-Moore L., Wicken A. J., Shockman G. D. Effect of lipoteichoic acid and lipids on lysis of intact cells of Streptococcus faecalis. J Bacteriol. 1976 Sep;127(3):1582–1584. doi: 10.1128/jb.127.3.1582-1584.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cleveland R. F., Holtje J. V., Wicken A. J., Tomasz A., Daneo-Moore L., Shockman G. D. Inhibition of bacterial wall lysins by lipoteichoic acids and related compounds. Biochem Biophys Res Commun. 1975 Dec 1;67(3):1128–1135. doi: 10.1016/0006-291x(75)90791-3. [DOI] [PubMed] [Google Scholar]
  9. Cleveland R. F., Wicken A. J., Daneo-Moore L., Shockman G. D. Inhibition of wall autolysis in Streptococcus faecalis by lipoteichoic acid and lipids. J Bacteriol. 1976 Apr;126(1):192–197. doi: 10.1128/jb.126.1.192-197.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Coley J., Duckworth M., Baddiley J. Extraction and purification of lipoteichoic acids from Gram-positive bacteria. Carbohydr Res. 1975 Mar;40(1):41–52. doi: 10.1016/s0008-6215(00)82667-1. [DOI] [PubMed] [Google Scholar]
  11. Coley J., Duckworth M., Baddiley J. The occurrence of lipoteichoic acids in the membranes of gram-positive bacteria. J Gen Microbiol. 1972 Dec;73(3):587–591. doi: 10.1099/00221287-73-3-587. [DOI] [PubMed] [Google Scholar]
  12. Cornett J. B., Shockman G. D. Cellular lysis of Streptococcus faecalis induced with triton X-100. J Bacteriol. 1978 Jul;135(1):153–160. doi: 10.1128/jb.135.1.153-160.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dulley J. R., Grieve P. A. A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Anal Biochem. 1975 Mar;64(1):136–141. doi: 10.1016/0003-2697(75)90415-7. [DOI] [PubMed] [Google Scholar]
  14. Emdur L. I., Chiu T. H. Turnover of phosphatidylglycerol in Streptococcus sanguis. Biochem Biophys Res Commun. 1974 Aug 5;59(3):1137–1144. doi: 10.1016/s0006-291x(74)80097-5. [DOI] [PubMed] [Google Scholar]
  15. Eytan G. D. Use of liposomes for reconstitution of biological functions. Biochim Biophys Acta. 1982 Oct 20;694(2):185–202. doi: 10.1016/0304-4157(82)90024-7. [DOI] [PubMed] [Google Scholar]
  16. Glaser L., Lindsay B. The synthesis of lipoteichoic acid carrier. Biochem Biophys Res Commun. 1974 Aug 5;59(3):1131–1136. doi: 10.1016/s0006-291x(74)80096-3. [DOI] [PubMed] [Google Scholar]
  17. Helenius A., Simons K. Solubilization of membranes by detergents. Biochim Biophys Acta. 1975 Mar 25;415(1):29–79. doi: 10.1016/0304-4157(75)90016-7. [DOI] [PubMed] [Google Scholar]
  18. Hinks E. T., Daneo-Moore L., Braverman S. Effects of temperature on the autolytic enzyme system of Streptococcus faecalis. J Bacteriol. 1978 Nov;136(2):491–496. doi: 10.1128/jb.136.2.491-496.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Horne D., Hakenbeck R., Tomasz A. Secretion of lipids induced by inhibition of peptidoglycan synthesis in streptococci. J Bacteriol. 1977 Nov;132(2):704–717. doi: 10.1128/jb.132.2.704-717.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Horne D., Tomasz A. Release of lipoteichoic acid from Streptococcus sanguis: stimulation of release during penicillin treatment. J Bacteriol. 1979 Mar;137(3):1180–1184. doi: 10.1128/jb.137.3.1180-1184.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ingram L. O. Adaptation of membrane lipids to alcohols. J Bacteriol. 1976 Feb;125(2):670–678. doi: 10.1128/jb.125.2.670-678.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Inoue K., Kitagawa T. Effect of lipid composition on sensitivity of lipid membranes to Triton X-100. Biochim Biophys Acta. 1976 Feb 19;426(1):1–16. doi: 10.1016/0005-2736(76)90424-7. [DOI] [PubMed] [Google Scholar]
  23. Jolliffe L. K., Doyle R. J., Streips U. N. The energized membrane and cellular autolysis in Bacillus subtilis. Cell. 1981 Sep;25(3):753–763. doi: 10.1016/0092-8674(81)90183-5. [DOI] [PubMed] [Google Scholar]
  24. Joseph R., Shockman G. D. Synthesis and excretion of glycerol teichoic acid during growth of two streptococcal species. Infect Immun. 1975 Aug;12(2):333–338. doi: 10.1128/iai.12.2.333-338.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kessler R. E., Shockman G. D. Enzymatic deacylation of lipoteichoic acid by protoplasts of Streptococcus faecium (Streptococcus faecalis ATCC 9790). J Bacteriol. 1979 Mar;137(3):1176–1179. doi: 10.1128/jb.137.3.1176-1179.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kessler R. E., Shockman G. D. Precursor-product relationship of intracellular and extracellular lipoteichoic acids of Streptococcus faecium. J Bacteriol. 1979 Feb;137(2):869–877. doi: 10.1128/jb.137.2.869-877.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kessler R. E., van de Rijn I., McCarty M. Characterization and localization of the enzymatic deacylation of lipoteichoic acid in group A streptococci. J Exp Med. 1979 Dec 1;150(6):1498–1509. doi: 10.1084/jem.150.6.1498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Koch H. U., Haas R., Fischer W. The role of lipoteichoic acid biosynthesis in membrane lipid metabolism of growing Staphylococcus aureus. Eur J Biochem. 1984 Jan 16;138(2):357–363. doi: 10.1111/j.1432-1033.1984.tb07923.x. [DOI] [PubMed] [Google Scholar]
  29. Komor E., Weber H., Tanner W. Greatly decreased susceptibility of nonmetabolizing cells towards detergents. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1814–1818. doi: 10.1073/pnas.76.4.1814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Koyama T., Yamada M., Matsuhashi M. Formation of regular packets of Staphylococcus aureus cells. J Bacteriol. 1977 Mar;129(3):1518–1523. doi: 10.1128/jb.129.3.1518-1523.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Lacks S. Mutants of Diplococcus pneumoniae that lack deoxyribonucleases and other activities possibly pertinent to genetic transformation. J Bacteriol. 1970 Feb;101(2):373–383. doi: 10.1128/jb.101.2.373-383.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lanyi J. K. Influence of electron transport on the interaction between membrane lipids and Triton X-100 in Halobacterium cutirubrum. Biochemistry. 1973 Mar 27;12(7):1433–1438. doi: 10.1021/bi00731a025. [DOI] [PubMed] [Google Scholar]
  34. Markham J. L., Knox K. W., Wicken A. J., Hewett M. J. Formation of extracellular lipoteichoic acid by oral streptococci and lactobacilli. Infect Immun. 1975 Aug;12(2):378–386. doi: 10.1128/iai.12.2.378-386.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Miozzari G. F., Niederberger P., Hütter R. Permeabilization of microorganisms by Triton X-100. Anal Biochem. 1978 Oct 1;90(1):220–233. doi: 10.1016/0003-2697(78)90026-x. [DOI] [PubMed] [Google Scholar]
  36. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  37. PARK J. T., JOHNSON M. J. A submicrodetermination of glucose. J Biol Chem. 1949 Nov;181(1):149–151. [PubMed] [Google Scholar]
  38. PATTERSON M. S., GREENE R. C. MEASUREMENT OF LOW ENERGY BETA-EMITTERS IN AQUEOUS SOLUTION BY LIQUID SCINTILLATION COUNTING OF EMULSIONS. Anal Chem. 1965 Jun;37:854–857. doi: 10.1021/ac60226a017. [DOI] [PubMed] [Google Scholar]
  39. Razin S. Reconstruction of biological membranes. Biochim Biophys Acta. 1972 Apr 18;265(2):241–296. [PubMed] [Google Scholar]
  40. Robinson N. C., Capaldi R. A. Interaction of detergents with cytochrome c oxidase. Biochemistry. 1977 Feb 8;16(3):375–381. doi: 10.1021/bi00622a006. [DOI] [PubMed] [Google Scholar]
  41. Schnaitman C. A. Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100. J Bacteriol. 1971 Oct;108(1):545–552. doi: 10.1128/jb.108.1.545-552.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sharon N., Jeanloz R. W. A procedure for the preparation of gram-quantities of bacterial cell walls. Experientia. 1964 May 15;20(5):253–254. doi: 10.1007/BF02151786. [DOI] [PubMed] [Google Scholar]
  43. Short S. A., White D. C., Kaback H. R. Active transport in isolated bacterial membrane vesicles. V. The transport of amino acids by membrane vesicles prepared from Staphylococcus aureus. J Biol Chem. 1972 Jan 10;247(1):298–304. [PubMed] [Google Scholar]
  44. Takebe I., Singer H. J., Wise E. M., Jr, Park J. T. Staphylococcus aureus H autolytic activity: general properties. J Bacteriol. 1970 Apr;102(1):14–19. doi: 10.1128/jb.102.1.14-19.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tanford C., Reynolds J. A. Characterization of membrane proteins in detergent solutions. Biochim Biophys Acta. 1976 Oct 26;457(2):133–170. doi: 10.1016/0304-4157(76)90009-5. [DOI] [PubMed] [Google Scholar]
  46. Tilby M. J. Detergent-resistant variants of Bacillus subtilis with reduced cell diameter. J Bacteriol. 1978 Oct;136(1):10–18. doi: 10.1128/jb.136.1.10-18.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tomasz A., Waks S. Mechanism of action of penicillin: triggering of the pneumococcal autolytic enzyme by inhibitors of cell wall synthesis. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4162–4166. doi: 10.1073/pnas.72.10.4162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Umbreit J. N., Strominger J. L. Relation of detergent HLB number to solubilization and stabilization of D-alanine carboxypeptidase from Bacillus subtilis membranes. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2997–3001. doi: 10.1073/pnas.70.10.2997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Ved H. S., Gustow E., Mahadevan V., Pieringer R. A. Dodecylglycerol. A new type of antibacterial agent which stimulates autolysin activity in Streptococcus faecium ATCC 9790. J Biol Chem. 1984 Jul 10;259(13):8115–8121. [PubMed] [Google Scholar]
  50. White D. C., Frerman F. E. Extraction, characterization, and cellular localization of the lipids of Staphylococcus aureus. J Bacteriol. 1967 Dec;94(6):1854–1867. doi: 10.1128/jb.94.6.1854-1867.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wicken A. J., Gibbens J. W., Knox K. W. Comparative studies on the isolation of membrane lipoteichoic acid from Lactobacillus fermenti. J Bacteriol. 1973 Jan;113(1):365–372. doi: 10.1128/jb.113.1.365-372.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wicken A. J., Knox K. W. Bacterial cell surface amphiphiles. Biochim Biophys Acta. 1980 May 27;604(1):1–26. doi: 10.1016/0005-2736(80)90583-0. [DOI] [PubMed] [Google Scholar]
  53. Wilkinson S. G. Glycosyl diglycerides from Pseudomonas rubescens. Biochim Biophys Acta. 1968 Oct 22;164(2):148–156. doi: 10.1016/0005-2760(68)90141-0. [DOI] [PubMed] [Google Scholar]
  54. Williamson R., Ward J. B. Deficiency of autolytic activity in Bacillus subtilis and Streptococcus pneumoniae is associated with a decreased permeability of the wall. J Gen Microbiol. 1981 Aug;125(2):325–334. doi: 10.1099/00221287-125-2-325. [DOI] [PubMed] [Google Scholar]
  55. Wong W., Chatterjee A. N., Young F. E. Regulation of bacterial cell walls: correlation between autolytic activity and cell wall turnover in Staphylococcus aureus. J Bacteriol. 1978 May;134(2):555–561. doi: 10.1128/jb.134.2.555-561.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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