Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1976 Aug;127(2):874–880. doi: 10.1128/jb.127.2.874-880.1976

Phospholipid composition and phospholipase A activity of Neisseria gonorrhoeae.

L M Senff, W S Wegener, G F Brooks, W R Finnerty, R A Makula
PMCID: PMC232996  PMID: 821921

Abstract

Exponential-phase cells of Neisseria gonorrhaeae 2686 were examined for phospholipid composition and for membrane-associated phospholipase A activity. When cells were harvested by centrifugation, washed, and lyophilized before extraction, approximately 74% of the total phospholipid was phosphatidylethanolamine, 18% was phosphatidylglycerol, 2% was cardiolipin, and 10% was lysophosphatidylethanolamine. However, when cells still suspended in growth medium were extracted, the amount of lysophosphatidylethanolamine decreased to approximately 1% of the phospholipid composition. This suggests that a gonococcal phospholipase A may be activated by conditions encountered during centrifugation and/or lyophilization of cells preceding extraction. Phospholipase A activity associated with cell membranes was assayed by measuring the conversion of tritiated phosphatidylethanolamine to lysophosphatidylethanolamine. Optimal activity was demonstrated in 10% methanol at pH 8.0 to 8.5, in the presence of calcium ions. The activity was both detergent sensitive and thermolabile. Comparisons of gonococcal colony types 1 and 4 showed no significant differences between the two types with respect to either phospholipid content or phospholipase A activity.

Full text

PDF
880

Images in this article

Selected References

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

  1. Albright F. R., White D. A., Lennarz W. J. Studies on enzymes involved in the catabolism of phospholipids in Escherichia coli. J Biol Chem. 1973 Jun 10;248(11):3968–3977. [PubMed] [Google Scholar]
  2. Audet A., Nantel G., Proulx P. Phospholipase A activity in growing Escherichia coli cells. Biochim Biophys Acta. 1974 Jun 26;348(3):334–343. doi: 10.1016/0005-2760(74)90213-6. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Barbu E., Lux M. Transformation des phospholipides bactériens conséctive à l'action du complément. C R Acad Sci Hebd Seances Acad Sci D. 1969 Jan 13;268(2):449–452. [PubMed] [Google Scholar]
  5. Brookes R., Hedén C. G. Dense cultures of Neisseria gonorrhoeae in liquid medium. Appl Microbiol. 1967 Mar;15(2):219–223. doi: 10.1128/am.15.2.219-223.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brooks J. B., Kellogg D. S., Thacker L., Turner E. M. Analysis by gas chromatography of fatty acids found in whole cultural extracts of Neisseria species. Can J Microbiol. 1971 Apr;17(4):531–543. doi: 10.1139/m71-088. [DOI] [PubMed] [Google Scholar]
  7. DITTMER J. C., LESTER R. L. A SIMPLE, SPECIFIC SPRAY FOR THE DETECTION OF PHOSPHOLIPIDS ON THIN-LAYER CHROMATOGRAMS. J Lipid Res. 1964 Jan;5:126–127. [PubMed] [Google Scholar]
  8. Doi O., Oki M., Nojima S. Two kinds of phospholipase A and lysophospholipase in Escherichia coli. Biochim Biophys Acta. 1972 Feb 21;260(2):244–258. [PubMed] [Google Scholar]
  9. Drach G., Cavard D., Barbu E. Transformation des phospholipides des formes L du streptocoque du groupe A sous l'action du complément du sérum humain. C R Acad Sci Hebd Seances Acad Sci D. 1973 Nov 12;277(19):2085–2088. [PubMed] [Google Scholar]
  10. Goldfine H. Comparative aspects of bacterial lipids. Adv Microb Physiol. 1972;8:1–58. doi: 10.1016/s0065-2911(08)60187-3. [DOI] [PubMed] [Google Scholar]
  11. Graham A. B., Wood G. C. On the activation of microsomal UDPglucuronyltransferase by phospholipase A. Biochim Biophys Acta. 1974 Dec 29;370(2):431–440. doi: 10.1016/0005-2744(74)90104-1. [DOI] [PubMed] [Google Scholar]
  12. Hebeler B. H., Young F. E. Autolysis of Neisseria gonorrhoeae. J Bacteriol. 1975 May;122(2):385–392. doi: 10.1128/jb.122.2.385-392.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. James-Holmquest A. N., Swanson J., Buchanan T. M., Wende R. D., Williams R. P. Differential attachment by piliated and nonpiliated Neisseria gonorrhoeae to human sperm. Infect Immun. 1974 May;9(5):897–902. doi: 10.1128/iai.9.5.897-902.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kent C., Lennarz W. J. An osmotically fragile mutant of Bacillus subtilis with an active membrane-associated phospholipase A 1 . Proc Natl Acad Sci U S A. 1972 Oct;69(10):2793–2797. doi: 10.1073/pnas.69.10.2793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Krag S. S., Lennarz W. J. Purification and characterization of an inhibitor of phospholipase A1 in Bacillus subtilis. J Biol Chem. 1975 Apr 25;250(8):2813–2822. [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. Ley H. L., Jr, Mueller J. H. On the Isolation from Agar of an Inhibitor for Neisseria gonorrhoeae. J Bacteriol. 1946 Oct;52(4):453–460. doi: 10.1128/jb.52.4.453-460.1946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Makula R. A., Finnerty W. R. Microbial assimilation of hydrocarbons: phospholipid metabolism. J Bacteriol. 1971 Sep;107(3):806–814. doi: 10.1128/jb.107.3.806-814.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Makula R. A., Finnerty W. R. Phospholipid composition of Desulfovibrio species. J Bacteriol. 1974 Dec;120(3):1279–1283. doi: 10.1128/jb.120.3.1279-1283.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mayer L. W., Holmes K. K., Falkow S. Characterization of plasmid deoxyribonucleic acid from Neisseria gonorrhoeae. Infect Immun. 1974 Oct;10(4):712–717. doi: 10.1128/iai.10.4.712-717.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mookerjea S., Yung J. W. A study on the effect of lysolecithin and phospholipase A on membrane-bound galactosyltransferase. Can J Biochem. 1974 Nov;52(11):1053–1066. doi: 10.1139/o74-147. [DOI] [PubMed] [Google Scholar]
  22. Morse S. A., Bartenstein L. Factors affecting autolysis of Neisseria gonorrhoeae. Proc Soc Exp Biol Med. 1974 Apr;145(4):1418–1421. doi: 10.3181/00379727-145-38025. [DOI] [PubMed] [Google Scholar]
  23. Ono Y., Nojima S. Phospholipases of the membrane fraction of Mycobacterium phlei. Biochim Biophys Acta. 1969 Jan 21;176(1):111–119. [PubMed] [Google Scholar]
  24. Punsalang A. P., Jr, Sawyer W. D. Role of pili in the virulence of Neisseria gonorrhoeae. Infect Immun. 1973 Aug;8(2):255–263. doi: 10.1128/iai.8.2.255-263.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Robinson J. M., Wilkinson J. H. The effect of phospholipases on the release of enzymes from intact cells. Clin Chim Acta. 1973 Sep 14;47(3):347–356. doi: 10.1016/0009-8981(73)90266-0. [DOI] [PubMed] [Google Scholar]
  26. SNYDER F., STEPHENS N. A simplified spectrophotometric determination of ester groups in lipids. Biochim Biophys Acta. 1959 Jul;34:244–245. doi: 10.1016/0006-3002(59)90255-0. [DOI] [PubMed] [Google Scholar]
  27. Scandella C. J., Kornberg A. A membrane-bound phospholipase A1 purified from Escherichia coli. Biochemistry. 1971 Nov 23;10(24):4447–4456. doi: 10.1021/bi00800a015. [DOI] [PubMed] [Google Scholar]
  28. Sud I. J., Feingold D. S. Phospholipids and fatty acids of Neisseria gonorrhoeae. J Bacteriol. 1975 Nov;124(2):713–717. doi: 10.1128/jb.124.2.713-717.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Swanson J. Studies on gonococcus infection. IV. Pili: their role in attachment of gonococci to tissue culture cells. J Exp Med. 1973 Mar 1;137(3):571–589. doi: 10.1084/jem.137.3.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Thongthai C., Sawyer W. D. Studies on the virulence of Neisseria gonorrhoeae. I. Relation of colonial morphology and resistance to phagocytosis by polymorphonuclear leukocytes. Infect Immun. 1973 Mar;7(3):373–379. doi: 10.1128/iai.7.3.373-379.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Walstad D. L., Reitz R. C., Sparling P. F. Growth inhibition among strains of Neisseria gonorrhoeae due to production of inhibitory free fatty acids and lysophosphatidylethanolamine: absence of bacteriocins. Infect Immun. 1974 Sep;10(3):481–488. doi: 10.1128/iai.10.3.481-488.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ward M. E., Watt P. J., Robertson J. N. The human fallopian tube: a laboratory model for gonococcal infection. J Infect Dis. 1974 Jun;129(6):650–659. doi: 10.1093/infdis/129.6.650. [DOI] [PubMed] [Google Scholar]
  33. Wolf-Watz H., Elmros T., Normark S., Bloom G. D. Cell envelope of Neisseria gonorrhoeae: outer membrane and peptidoglycan composition of penicillin-sensitive and-resistant strains. Infect Immun. 1975 Jun;11(6):1332–1341. doi: 10.1128/iai.11.6.1332-1341.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. de Haas G. H., Sarda L., Roger J. Positional specific hydrolysis of phospholipids by pancreatic lipase. Biochim Biophys Acta. 1965 Dec 2;106(3):638–640. doi: 10.1016/0005-2760(65)90082-2. [DOI] [PubMed] [Google Scholar]

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

RESOURCES