Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Aug;151(2):629–635. doi: 10.1128/jb.151.2.629-635.1982

High-density lipoprotein that supports Ureaplasma urealyticum growth.

I A Sayed, F W Sweat
PMCID: PMC220303  PMID: 7201468

Abstract

A high-density lipoprotein with growth-promoting activity for Ureaplasma urealyticum was purified in high yield from equine serum by ammonium sulfate fractionation and molecular filtration. Fractions enriched in growth-promoting activity represented 5% of the total serum protein, and 30 micrograms of the purified protein per ml gave an activity equivalent to that from 100 micrograms of whole serum per ml. The serum was totally replaced by purified lipoprotein when tested in a soy peptone-yeast dialysate or when added to a chemically defined synthetic medium. Polyacrylamide gel electrophoresis indicated that one major protein with growth-promoting activity is present. A total of 10 proteins were distinguished by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with 75% of the total contributed by two proteins with molecular weights of 160,000 and 170,000. A total of 90% of the lipoprotein was an alpha-protein with a mobility of 0.67 in two-dimensional immunoelectrophoresis (albumin = 1.0). The active component was further characterized as high-density lipoprotein by density ultracentrifugation. Two components with S = 6.4 and S = 15.8 were distinguished by velocity sedimentation. The lipid was removed from lipoprotein during its precipitation with acetone. The growth-promoting activity of delipidized protein was dependent upon the addition of exogenous cholesterol, and [14C]cholesterol was transferred to urea-plasmic cells in cultures containing the delipidized protein. A major portion of the [14C]cholesterol remained associated with the protein during filtration on Sepharose 4B columns.

Full text

PDF
629

Images in this article

633Image
on p.633

Selected References

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

  1. 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]
  2. Bowden J. A., Fried M. Separation and characterization of rat plasma lipoproteins by molecular sieve chromatography. Comp Biochem Physiol. 1970 Feb 1;32(3):391–400. doi: 10.1016/0010-406x(70)90456-1. [DOI] [PubMed] [Google Scholar]
  3. EDWARD D. G., FITZGERALD W. A. Cholesterol in the growth of organisms of the pleuropneumonia group. J Gen Microbiol. 1951 Aug;5(3):576–586. doi: 10.1099/00221287-5-3-576. [DOI] [PubMed] [Google Scholar]
  4. Fainaru M., Havel R. J., Imaizumi K. Apoprotein content of plasma lipoproteins of the rat separated by gel chromatography or ultracentrifugation. Biochem Med. 1977 Jun;17(3):347–355. doi: 10.1016/0006-2944(77)90040-0. [DOI] [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. HAVEL R. J., EDER H. A., BRAGDON J. H. The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum. J Clin Invest. 1955 Sep;34(9):1345–1353. doi: 10.1172/JCI103182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harboe N., Ingild A. Immunization, isolation of immunoglobulins, estimation of antibody titre. Scand J Immunol Suppl. 1973;1:161–164. doi: 10.1111/j.1365-3083.1973.tb03798.x. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. 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]
  11. 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]
  12. Nicoll A., Miller N. E., Lewis B. High-density lipoprotein metabolism. Adv Lipid Res. 1980;17:53–106. doi: 10.1016/b978-0-12-024917-6.50008-2. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Romano N., Rottem S., Razin S. Biosynthesis of saturated and unsaturated fatty acids by a T-strain mycoplasma (Ureaplasma). J Bacteriol. 1976 Oct;128(1):170–173. doi: 10.1128/jb.128.1.170-173.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rottem S. Membrane lipids of mycoplasmas. Biochim Biophys Acta. 1980 May 27;604(1):65–90. doi: 10.1016/0005-2736(80)90585-4. [DOI] [PubMed] [Google Scholar]
  16. Rottem S., Pfendt E. A., Hayflick L. Sterol requirements of T-strain mycoplasmas. J Bacteriol. 1971 Jan;105(1):323–330. doi: 10.1128/jb.105.1.323-330.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. SMITH P. F., LECCE J. G., LYNN R. J. A lipoprotein as a growth factor for cortain pleuropneumonialike organisms. J Bacteriol. 1954 Nov;68(5):627–633. doi: 10.1128/jb.68.5.627-633.1954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. SMITH P. F., MORTON H. E. Further characterization of the protein factor required by certain pleuropneumonia-like organisms for growth in vitro. Arch Biochem Biophys. 1952 Jul;38:23–28. doi: 10.1016/0003-9861(52)90004-0. [DOI] [PubMed] [Google Scholar]
  19. SMITH P. F., MORTON H. E. The separation and characterization of the growth factor in serum and ascitic fluid which is required by certain pleuropneumonia-like organisms. J Bacteriol. 1951 Apr;61(4):395–405. doi: 10.1128/jb.61.4.395-405.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sayed I. A., Kenny G. E. Effect of ammonium ion concentration and osmotic pressure on growth of Ureaplasma urealyticum (T-strain mycoplasma). J Bacteriol. 1978 Jun;134(3):967–972. doi: 10.1128/jb.134.3.967-972.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sayed I. A., Kenny G. E. Effect of reducing agents, catalase, and reuse of medium on toxicity of media for growth of Ureaplasma urealyticum. J Infect Dis. 1979 Jun;139(6):720–724. doi: 10.1093/infdis/139.6.720. [DOI] [PubMed] [Google Scholar]
  22. Slutzky G. M., Razin S., Kahane I., Eisenberg S. Cholesterol transfer from serum lipoproteins to mycoplasma membranes. Biochemistry. 1977 Nov 15;16(23):5158–5163. doi: 10.1021/bi00642a032. [DOI] [PubMed] [Google Scholar]
  23. Slutzky G. M., Razin S., Kahane I. Serum lipoproteins as cholesterol donors to mycoplasma membranes. Biochem Biophys Res Commun. 1976 Jan 26;68(2):529–536. doi: 10.1016/0006-291x(76)91178-5. [DOI] [PubMed] [Google Scholar]
  24. Smith P. F., Rothblat G. H. INCORPORATION OF CHOLESTEROL BY PLEUROPNEUMONIA-LIKE ORGANISMS. J Bacteriol. 1960 Dec;80(6):842–850. doi: 10.1128/jb.80.6.842-850.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sodhi H. S., Gould R. G. Combination of delipidized high density lipoprotein with lipids. J Biol Chem. 1967 Mar 25;242(6):1205–1210. [PubMed] [Google Scholar]
  26. Tall A. R., Deckelbaum R. J., Small D. M., Shipley G. G. Thermal behavior of human plasma high density lipoprotein. Biochim Biophys Acta. 1977 Apr 26;487(1):145–133. doi: 10.1016/0005-2760(77)90051-0. [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. Vitello L. B., Scanu A. M. Studies on human serum high-density lipoproteins. Self-association of human serum apolipoprotein A-II in aqueous solutions. Biochemistry. 1976 Mar 9;15(5):1161–1165. doi: 10.1021/bi00650a031. [DOI] [PubMed] [Google Scholar]
  29. Washburn L. R., Hughes J. H., Somerson N. L. Mycoplasma growth factors in bovine serum fraction. J Bacteriol. 1978 Sep;135(3):818–827. doi: 10.1128/jb.135.3.818-827.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yaguzhinskaya O. E. Detection of serum proteins in the electrophoretic patterns of total proteins of mycoplasma cells. J Hyg (Lond) 1976 Oct;77(2):189–198. doi: 10.1017/s002217240002461x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES