Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Jun;170(6):2692–2697. doi: 10.1128/jb.170.6.2692-2697.1988

Characteristics of Ureaplasma urealyticum urease.

A Blanchard 1, S Razin 1, G E Kenny 1, M F Barile 1
PMCID: PMC211190  PMID: 3131306

Abstract

Sonication of Ureaplasma urealyticum cells grown in a dialysate growth medium effectively separated the cytoplasmic fraction from the membrane fraction, with both fractions relatively free from exogenous contaminating proteins. The urease activity was associated with the cytoplasmic fraction, and the ureaplasmal urease exhibited a specific activity higher than that of crystalline jack bean urease. The enzymatic activity of the ureaplasmal enzyme was optimum at pH 7.5 and was resistant to the chelating agents EDTA and sodium citrate. Sulfhydryl-blocking agents such as HgCl2 and Pb(NO3)2 inhibited the ureaplasmal urease, which was also shown to be particularly sensitive to flurofamide and, to a much lesser extent, to acetohydroxamic acid. Electrophoretic analysis of the proteins of the ureaplasmal cell fractions combined with Western immunoblot with an antiserum to the ureaplasmal urease indicated that the urease constitutes a major component of the cytoplasm and is composed of several 70-kilodalton polypeptides.

Full text

PDF
2696

Images in this article

2695Image
on p.2695
2695Image
on p.2695
2695Image
on p.2695

Selected References

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

  1. Alexander A. G., Kenny G. E. Characterization of membrane and cytoplasmic antigens of Mycoplasma arginini by two-dimensional (crossed) immunoelectrophoresis. Infect Immun. 1977 Jan;15(1):313–321. doi: 10.1128/iai.15.1.313-321.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dixon N. E., Gazzola T. C., blakeley R. L., Zermer B. Letter: Jack bean urease (EC 3.5.1.5). A metalloenzyme. A simple biological role for nickel? J Am Chem Soc. 1975 Jul 9;97(14):4131–4133. doi: 10.1021/ja00847a045. [DOI] [PubMed] [Google Scholar]
  3. Eng H., Robertson J. A., Stemke G. W. Properties of urease from Ureaplasma urealyticum: kinetics, molecular weight, and demonstration of multiple enzyme isoelectric point forms. Can J Microbiol. 1986 Jun;32(6):487–493. doi: 10.1139/m86-089. [DOI] [PubMed] [Google Scholar]
  4. Ford D. K. Inhibition of growth of T-strain mycoplasmas by hydroxamic acids and by aurothiomalate. Antimicrob Agents Chemother. 1972 Nov;2(5):340–343. doi: 10.1128/aac.2.5.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ford D. K., MacDonald J. Influence of urea on the growth of T-strain mycoplasmas. J Bacteriol. 1967 May;93(5):1509–1512. doi: 10.1128/jb.93.5.1509-1512.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kenny G. E., Cartwright F. D. Effect of urea concentration on growth of Ureaplasma urealyticum (T-strain mycoplasma). J Bacteriol. 1977 Oct;132(1):144–150. doi: 10.1128/jb.132.1.144-150.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kenny G. E. Heat-lability and organic solvent-solubility of mycoplasma antigens. Ann N Y Acad Sci. 1967 Jul 28;143(1):676–681. doi: 10.1111/j.1749-6632.1967.tb27713.x. [DOI] [PubMed] [Google Scholar]
  8. Kenny G. E. Inhibition of the growth of Ureaplasma urealyticum by a new urease inhibitor, flurofamide. Yale J Biol Med. 1983 Sep-Dec;56(5-6):717–722. [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Masover G. K., Razin S., Hayflick L. Effects of carbon dioxide, urea, and ammonia on growth of Ureaplasma urealyticum (T-strain mycoplasma). J Bacteriol. 1977 Apr;130(1):292–296. doi: 10.1128/jb.130.1.292-296.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Masover G. K., Razin S., Hayflick L. Localization of enzymes in Ureaplasma urealyticum (T-strain mycoplasma). J Bacteriol. 1977 Apr;130(1):297–302. doi: 10.1128/jb.130.1.297-302.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Masover G. K., Sawyer J. E., Hayflick L. Urea-hydrolyzing activity of a T-strain mycoplasma: Ureaplasma urealyticum. J Bacteriol. 1976 Feb;125(2):581–587. doi: 10.1128/jb.125.2.581-587.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Millner O. E., Jr, Andersen J. A., Appler M. E., Benjamin C. E., Edwards J. G., Humphrey D. T., Shearer E. M. Flurofamide: a potent inhibitor of bacterial urease with potential clinical utility in the treatment of infection induced urinary stones. J Urol. 1982 Feb;127(2):346–350. doi: 10.1016/s0022-5347(17)53779-9. [DOI] [PubMed] [Google Scholar]
  15. Mobley H. L., Jones B. D., Jerse A. E. Cloning of urease gene sequences from Providencia stuartii. Infect Immun. 1986 Oct;54(1):161–169. doi: 10.1128/iai.54.1.161-169.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pollack J. D. Metabolic distinctiveness of ureaplasmas. Pediatr Infect Dis. 1986 Nov-Dec;5(6 Suppl):S305–S307. doi: 10.1097/00006454-198611010-00023. [DOI] [PubMed] [Google Scholar]
  17. Precious B. L., Thirkell D., Russell W. C. Preliminary characterization of the urease and a 96 kDa surface-expressed polypeptide of Ureaplasma urealyticum. J Gen Microbiol. 1987 Sep;133(9):2659–2670. doi: 10.1099/00221287-133-9-2659. [DOI] [PubMed] [Google Scholar]
  18. RAZIN S., ARGAMAN M., AVIGAN J. CHEMICAL COMPOSITION OF MYCOPLASMA CELLS AND MEMBRANES. J Gen Microbiol. 1963 Dec;33:477–487. doi: 10.1099/00221287-33-3-477. [DOI] [PubMed] [Google Scholar]
  19. Razin S. The mycoplasmas. Microbiol Rev. 1978 Jun;42(2):414–470. doi: 10.1128/mr.42.2.414-470.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Romano N., La Licata R. Cell fractions and enzymatic activities of Ureaplasma urealyticum. J Bacteriol. 1978 Dec;136(3):833–838. doi: 10.1128/jb.136.3.833-838.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Romano N., La Licata R., Russo Alesi D. Energy production in Ureaplasma urealyticum. Pediatr Infect Dis. 1986 Nov-Dec;5(6 Suppl):S308–S312. doi: 10.1097/00006454-198611010-00024. [DOI] [PubMed] [Google Scholar]
  22. Romano N., Tolone G., Ajello F., La Licata R. Adenosine 5'-triphosphate synthesis induced by urea hydrolysis in Ureaplasma urealyticum. J Bacteriol. 1980 Nov;144(2):830–832. doi: 10.1128/jb.144.2.830-832.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Senior B. W., Bradford N. C., Simpson D. S. The ureases of Proteus strains in relation to virulence for the urinary tract. J Med Microbiol. 1980 Nov;13(4):507–512. doi: 10.1099/00222615-13-4-507. [DOI] [PubMed] [Google Scholar]
  24. Shepard M. C., Lunceford C. D. Occurrence of urease in T strains of Mycoplasma. J Bacteriol. 1967 May;93(5):1513–1520. doi: 10.1128/jb.93.5.1513-1520.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stemke G. W., Robertson J. A., Nhan M. Purification of urease from Ureaplasma urealyticum. Can J Microbiol. 1987 Oct;33(10):857–862. doi: 10.1139/m87-150. [DOI] [PubMed] [Google Scholar]
  26. Swanberg S. L., Masover G. K., Hayflick L. Some characteristics of Ureaplasma urealyticum. Urease activity in a simple buffer: effect of metal ions and sulphydryl inhibitors. J Gen Microbiol. 1978 Oct;108(2):221–225. doi: 10.1099/00221287-108-2-221. [DOI] [PubMed] [Google Scholar]
  27. Thirkill C. E., Kenny G. E. Serological comparison of five arginine-utilizing Mycoplasma species by two-dimensional immunoelectrophoresis. Infect Immun. 1974 Sep;10(3):624–632. doi: 10.1128/iai.10.3.624-632.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES