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. 1983 Jun;17(6):1132–1140. doi: 10.1128/jcm.17.6.1132-1140.1983

Electrophoretic characterization of soluble protein extracts of Legionella pneumophila and other members of the family Legionellaceae.

M Lema, A Brown
PMCID: PMC272814  PMID: 6874904

Abstract

The soluble peptides of strains of Legionella pneumophila, Tatlockia micdadei, Fluoribacter bozemanae, Fluoribacter dumoffii, and Fluoribacter gormanii were studied by polyacrylamide gel electrophoresis. Characteristic patterns were seen for Legionella and Tatlockia strains, whereas the patterns for the Fluoribacter strains were variable as would be expected for this genetically heterogeneous group. Grouping by peptide pattern was consistent with proposed taxons based on DNA-DNA homology. By using a new silver stain technique, the sensitivity and ease of pattern recognition were enhanced significantly. This technique is an easily applied general method for distinguishing between strains in epidemiological studies.

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Selected References

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  1. Baine W. B., Rasheed J. K. Aromatic substrate specificity of browning by cultures of the Legionnaires' disease bacterium. Ann Intern Med. 1979 Apr;90(4):619–620. doi: 10.7326/0003-4819-90-4-619. [DOI] [PubMed] [Google Scholar]
  2. Berdal B. P., Hushovd O., Olsvik O., odegård O. R., Bergan T. Demonstration of extracellular proteolytic enzymes from Legionella species strains by using synthetic chromogenic peptide substrates. Acta Pathol Microbiol Immunol Scand B. 1982 Apr;90(2):119–123. doi: 10.1111/j.1699-0463.1982.tb00092.x. [DOI] [PubMed] [Google Scholar]
  3. Bibb W. F., Sorg R. J., Thomason B. M., Hicklin M. D., Steigerwalt A. G., Brenner D. J., Wulf M. R. Recognition of a second serogroup of Legionella longbeachae. J Clin Microbiol. 1981 Dec;14(6):674–677. doi: 10.1128/jcm.14.6.674-677.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brenner D. J., Steigerwalt A. G., McDade J. E. Classification of the Legionnaires' disease bacterium: Legionella pneumophila, genus novum, species nova, of the family Legionellaceae, familia nova. Ann Intern Med. 1979 Apr;90(4):656–658. doi: 10.7326/0003-4819-90-4-656. [DOI] [PubMed] [Google Scholar]
  5. Brown A., Vickers R. M., Elder E. M., Lema M., Garrity G. M. Plasmid and surface antigen markers of endemic and epidemic Legionella pneumophila strains. J Clin Microbiol. 1982 Aug;16(2):230–235. doi: 10.1128/jcm.16.2.230-235.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cato E. P., Hash D. E., Holdeman L. V., Moore W. E. Electrophoretic study of Clostridium species. J Clin Microbiol. 1982 Apr;15(4):688–702. doi: 10.1128/jcm.15.4.688-702.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cherry W. B., Gorman G. W., Orrison L. H., Moss C. W., Steigerwalt A. G., Wilkinson H. W., Johnson S. E., McKinney R. M., Brenner D. J. Legionella jordanis: a new species of Legionella isolated from water and sewage. J Clin Microbiol. 1982 Feb;15(2):290–297. doi: 10.1128/jcm.15.2.290-297.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cordes L. G., Wiesenthal A. M., Gorman G. W., Phair J. P., Sommers H. M., Brown A., Yu V. L., Magnussen M. H., Meyer R. D., Wolf J. S. Isolation of Legionella pneumophila from hospital shower heads. Ann Intern Med. 1981 Feb;94(2):195–197. doi: 10.7326/0003-4819-94-2-195. [DOI] [PubMed] [Google Scholar]
  9. Cordes L. G., Wilkinson H. W., Gorman G. W., Fikes B. J., Fraser D. W. Atypical Legionella-like organisms: fastidious water-associated bacteria pathogenic for man. Lancet. 1979 Nov 3;2(8149):927–930. doi: 10.1016/s0140-6736(79)92623-0. [DOI] [PubMed] [Google Scholar]
  10. Daniels M. J., Meddins B. M. Polyacrylamide gel electrophoresis of mycoplasma proteins in sodium dodecyl sulphate. J Gen Microbiol. 1973 May;76(1):239–242. doi: 10.1099/00221287-76-1-239. [DOI] [PubMed] [Google Scholar]
  11. Fliermans C. B., Cherry W. B., Orrison L. H., Smith S. J., Tison D. L., Pope D. H. Ecological distribution of Legionella pneumophila. Appl Environ Microbiol. 1981 Jan;41(1):9–16. doi: 10.1128/aem.41.1.9-16.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fliermans C. B., Cherry W. B., Orrison L. H., Thacker L. Isolation of Legionella pneumophila from nonepidemic-related aquatic habitats. Appl Environ Microbiol. 1979 Jun;37(6):1239–1242. doi: 10.1128/aem.37.6.1239-1242.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fraser D. W., Tsai T. R., Orenstein W., Parkin W. E., Beecham H. J., Sharrar R. G., Harris J., Mallison G. F., Martin S. M., McDade J. E. Legionnaires' disease: description of an epidemic of pneumonia. N Engl J Med. 1977 Dec 1;297(22):1189–1197. doi: 10.1056/NEJM197712012972201. [DOI] [PubMed] [Google Scholar]
  14. Garrity G. M., Elder E. M., Davis B., Vickers R. M., Brown A. Serological and genotypic diversity among serogroup 5- reacting environmental Legionella isolates. J Clin Microbiol. 1982 Apr;15(4):646–653. doi: 10.1128/jcm.15.4.646-653.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Glick T. H., Gregg M. B., Berman B., Mallison G., Rhodes W. W., Jr, Kassanoff I. Pontiac fever. An epidemic of unknown etiology in a health department: I. Clinical and epidemiologic aspects. Am J Epidemiol. 1978 Feb;107(2):149–160. doi: 10.1093/oxfordjournals.aje.a112517. [DOI] [PubMed] [Google Scholar]
  16. Gorman G. W., Yu V. L., Brown A., Hall J. A., Martin W. T., Bibb W. F., Morris G. K., Magnussen M. H., Fraser D. W. Isolation of Pittsburgh pneumonia agent from nebulizers used in respiratory therapy. Ann Intern Med. 1980 Oct;93(4):572–573. doi: 10.7326/0003-4819-93-4-572. [DOI] [PubMed] [Google Scholar]
  17. Hébert G. A. Hippurate hydrolysis by Legionella pneumophila. J Clin Microbiol. 1981 Jan;13(1):240–242. doi: 10.1128/jcm.13.1.240-242.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jarvis A. W., Wolff J. M. Grouping of lactic streptococci by gel electrophoresis of soluble cell extracts. Appl Environ Microbiol. 1979 Mar;37(3):391–398. doi: 10.1128/aem.37.3.391-398.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Karr D. E., Bibb W. F., Moss C. W. Isoprenoid quinones of the genus Legionella. J Clin Microbiol. 1982 Jun;15(6):1044–1048. doi: 10.1128/jcm.15.6.1044-1048.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kersters K., De Ley J. Identification and grouping of bacteria by numerical analysis of their electrophoretic protein patterns. J Gen Microbiol. 1975 Apr;87(2):333–342. doi: 10.1099/00221287-87-2-333. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Mayberry W. R. Dihydroxy and monohydroxy fatty acids in Legionella pneumophila. J Bacteriol. 1981 Aug;147(2):373–381. doi: 10.1128/jb.147.2.373-381.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McKinney R. M., Porschen R. K., Edelstein P. H., Bissett M. L., Harris P. P., Bondell S. P., Steigerwalt A. G., Weaver R. E., Ein M. E., Lindquist D. S. Legionella longbeachae species nova, another etiologic agent of human pneumonia. Ann Intern Med. 1981 Jun;94(6):739–743. doi: 10.7326/0003-4819-94-6-739. [DOI] [PubMed] [Google Scholar]
  25. McKinney R. M., Wilkinson H. W., Sommers H. M., Fikes B. J., Sasseville K. R., Yungbluth M. M., Wolf J. S. Legionella pneumophila serogroup six: isolation from cases of legionellosis, identification by immunofluorescence staining, and immunological response to infection. J Clin Microbiol. 1980 Sep;12(3):395–401. doi: 10.1128/jcm.12.3.395-401.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mocca L. F., Frasch C. E. Sodium dodecyl sulfate-polyacrylamide gel typing system for characterization of Neisseria meningitidis isolates. J Clin Microbiol. 1982 Aug;16(2):240–244. doi: 10.1128/jcm.16.2.240-244.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moore W. E., Hash D. E., Holdeman L. V., Cato E. P. Polyacrylamide slab gel electrophoresis of soluble proteins for studies of bacterial floras. Appl Environ Microbiol. 1980 Apr;39(4):900–907. doi: 10.1128/aem.39.4.900-907.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Morris G. K., Steigerwalt A., Feeley J. C., Wong E. S., Martin W. T., Patton C. M., Brenner D. J. Legionella gormanii sp. nov. J Clin Microbiol. 1980 Nov;12(5):718–721. doi: 10.1128/jcm.12.5.718-721.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Moss C. W., Karr D. E., Dees S. B. Cellular fatty acid composition of Legionella longbeachae sp. nov. J Clin Microbiol. 1981 Dec;14(6):692–694. doi: 10.1128/jcm.14.6.692-694.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Moss C. W., Weaver R. E., Dees S. B., Cherry W. B. Cellular fatty acid composition of isolates from Legionnaires disease. J Clin Microbiol. 1977 Aug;6(2):140–143. doi: 10.1128/jcm.6.2.140-143.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Myerowitz R. L., Pasculle A. W., Dowling J. N., Pazin G. J., Sr, Puerzer M., Yee R. B., Rinaldo C. R., Jr, Hakala T. R. Opportunistic lung infection due to "Pittsburgh Pneumonia Agent". N Engl J Med. 1979 Nov 1;301(18):953–958. doi: 10.1056/NEJM197911013011801. [DOI] [PubMed] [Google Scholar]
  32. Müller H. E. Enzymatic profile of Legionella pneumophilia. J Clin Microbiol. 1981 Mar;13(3):423–426. doi: 10.1128/jcm.13.3.423-426.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Noble R. C., Schell S. C. Acrylamide gel electrophoresis of proteins of Neisseria gonorrhoeae as an epidemiological tool. Infect Immun. 1978 Jan;19(1):178–186. doi: 10.1128/iai.19.1.178-186.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nolte F. S., Hollick G. E., Robertson R. G. Enzymatic activities of Legionella pneumophila and Legionella-like organisms. J Clin Microbiol. 1982 Jan;15(1):175–177. doi: 10.1128/jcm.15.1.175-177.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Orrison L. H., Cherry W. B., Fliermans C. B., Dees S. B., McDougal L. K., Dodd D. J. Characteristics of environmental isolates of Legionella pneumophila. Appl Environ Microbiol. 1981 Jul;42(1):109–115. doi: 10.1128/aem.42.1.109-115.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pasculle A. W., Feeley J. C., Gibson R. J., Cordes L. G., Myerowitz R. L., Patton C. M., Gorman G. W., Carmack C. L., Ezzell J. W., Dowling J. N. Pittsburgh pneumonia agent: direct isolation from human lung tissue. J Infect Dis. 1980 Jun;141(6):727–732. doi: 10.1093/infdis/141.6.727. [DOI] [PubMed] [Google Scholar]
  37. Rouatt J. W., Skyring G. W., Purkayastha V. Soil bacteria: numerical analysis of electrophoretic protein patterns developed in acrylamide gels. Can J Microbiol. 1970 Mar;16(3):202–205. doi: 10.1139/m70-035. [DOI] [PubMed] [Google Scholar]
  38. Swings J., De Ley J. The biology of Zymomonas. Bacteriol Rev. 1977 Mar;41(1):1–46. doi: 10.1128/br.41.1.1-46.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tobin J. O., Bartlett C. L., Waitkins S. A., Barrow G. I., Macrae A. D., Taylor A. G., Fallon R. J., Lynch F. R. Legionnaires' disease: further evidence to implicate water storage and distribution systems as sources. Br Med J (Clin Res Ed) 1981 Feb 14;282(6263):573–573. doi: 10.1136/bmj.282.6263.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Vickers R. M., Brown A., Garrity G. M. Dye-containing buffered charcoal-yeast extract medium for differentiation of members of the family Legionellaceae. J Clin Microbiol. 1981 Feb;13(2):380–382. doi: 10.1128/jcm.13.2.380-382.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wilkinson H. W., Fikes B. J., Cruce D. D. Indirect immunofluorescence test for serodiagnosis of Legionnaires disease: evidence for serogroup diversity of Legionnaires disease bacterial antigens and for multiple specificity of human antibodies. J Clin Microbiol. 1979 Mar;9(3):379–383. doi: 10.1128/jcm.9.3.379-383.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wilkinson H. W., Fikes B. J. Slide agglutination test for serogrouping Legionella pneumophila and atypical Legionella-like organisms. J Clin Microbiol. 1980 Jan;11(1):99–101. doi: 10.1128/jcm.11.1.99-101.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wong K. H., Schalla W. O., Arko R. J., Bullard J. C., Feeley J. C. Immunochemical, serologic, and immunologic properties of major antigens isolated from the Legionnaires' disease bacterium. Observations bearing on the feasibility of a vaccine. Ann Intern Med. 1979 Apr;90(4):634–638. doi: 10.7326/0003-4819-90-4-634. [DOI] [PubMed] [Google Scholar]

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