Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Oct;71(10):4072–4076. doi: 10.1073/pnas.71.10.4072

Manipulation of the Phospholipid Composition of Tissue Culture Cells*

Michael Glaser 1,†, Karen A Ferguson 1,‡, P Roy Vagelos 1
PMCID: PMC434330  PMID: 4530286

Abstract

Methods have been devised to significantly alter the phospholipid composition of LM cells grown in serum-free tissue culture medium. The polar head groups of the phospholipids, as well as the acyl groups of these lipids, can be changed. When LM cells were grown in medium containing choline analogues, either N-methyl-ethanolamine or N,N-dimethylethanolamine, or the unnatural analogues, 1-2-amino-1-butanol or 3-amino-1-propanol, up to 50% of the cellular phospholipids contained the analogue supplied. When linoleate was added to the cells as a bovine serum albumin complex, up to 40% of the fatty acids of the phospholipids were linoleate. Under the conditions discussed, the polar head group composition, the fatty acid composition, or both together could be varied in the membrane phospholipids.

Keywords: polar head group, fatty acid, LM cells, membrane

Full text

PDF
4072

Images in this article

4074Image
on p.4074

Selected References

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

  1. Ames G. F. Lipids of Salmonella typhimurium and Escherichia coli: structure and metabolism. J Bacteriol. 1968 Mar;95(3):833–843. doi: 10.1128/jb.95.3.833-843.1968. [DOI] [PMC free article] [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. CROCKEN B. J., NYC J. F. PHOSPHOLIPID VARIATIONS IN MUTANT STRAINS OF NEUROSPORA CRASSA. J Biol Chem. 1964 Jun;239:1727–1730. [PubMed] [Google Scholar]
  4. Cronan J. E., Vagelos P. R. Metabolism and function of the membrane phospholipids of Escherichia coli. Biochim Biophys Acta. 1972 Feb 14;265(1):25–60. doi: 10.1016/0304-4157(72)90018-4. [DOI] [PubMed] [Google Scholar]
  5. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dawson R. M. The formation of phosphatidylglycerol and other phospholipids by the transferase activity of phospholipase D. Biochem J. 1967 Jan;102(1):205–210. doi: 10.1042/bj1020205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Higuchi K. An improved chemically defined culture medium for strain L mouse cells based on growth responses to graded levels of nutrients including iron and zinc ions. J Cell Physiol. 1970 Feb;75(1):65–72. doi: 10.1002/jcp.1040750108. [DOI] [PubMed] [Google Scholar]
  8. Ladbrooke B. D., Chapman D. Thermal analysis of lipids, proteins and biological membranes. A review and summary of some recent studies. Chem Phys Lipids. 1969 Dec;3(4):304–356. doi: 10.1016/0009-3084(69)90040-1. [DOI] [PubMed] [Google Scholar]
  9. Lester R. L., White D. C. Quantitative gas-liquid chromatographic analysis of ethanolamine, monomethyl ethanolamine, and dimethyl ethanolamine from lipids. J Lipid Res. 1967 Nov;8(6):565–568. [PubMed] [Google Scholar]
  10. MUNKRES K. D., RICHARDS F. M. THE PURIFICATION AND PROPERTIES OF NEUROSPORA MALATE DEHYDROGENASE. Arch Biochem Biophys. 1965 Mar;109:466–479. doi: 10.1016/0003-9861(65)90391-7. [DOI] [PubMed] [Google Scholar]
  11. Machtiger N. A., Fox C. F. Biochemistry of bacterial membranes. Annu Rev Biochem. 1973;42:575–600. doi: 10.1146/annurev.bi.42.070173.003043. [DOI] [PubMed] [Google Scholar]
  12. Nagle S. C., Jr Improved growth of mammalian and insect cells in media containing increased levels of choline. Appl Microbiol. 1969 Feb;17(2):318–319. doi: 10.1128/am.17.2.318-319.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rittenhouse H. G., Fox C. F. Concanavalin A mediated hemagglutination and binding properties of LM cells. Biochem Biophys Res Commun. 1974 Mar 15;57(1):323–331. doi: 10.1016/s0006-291x(74)80393-1. [DOI] [PubMed] [Google Scholar]
  14. Rothblat G. H., Burns C. H., Conner R. L., Landrey J. R. Desmosterol as the major sterol in L-cell mouse fibroblasts grown in sterol-free culture medium. Science. 1970 Aug 28;169(3948):880–882. doi: 10.1126/science.169.3948.880. [DOI] [PubMed] [Google Scholar]
  15. Scarborough G. A., Nyc J. F. Methylation of ethanolamine phosphatides by microsomes from normal and mutant strains of Neurospora crassa. J Biol Chem. 1967 Jan 25;242(2):238–242. [PubMed] [Google Scholar]
  16. Sokoloff L., Rothblat G. H. Regulation of sterol synthesis in L cells: steady-state and transitional responses. Biochim Biophys Acta. 1972 Sep 7;280(1):172–181. doi: 10.1016/0005-2760(72)90222-6. [DOI] [PubMed] [Google Scholar]
  17. Spector A. A., Hoak J. C. An improved method for the addition of long-chain free fatty acid to protein solutions. Anal Biochem. 1969 Nov;32(2):297–302. doi: 10.1016/0003-2697(69)90089-x. [DOI] [PubMed] [Google Scholar]
  18. Vaskovsky V. E., Kostetsky E. Y. Modified spray for the detection of phospholipids on thin-layer chromatograms. J Lipid Res. 1968 May;9(3):396–396. [PubMed] [Google Scholar]
  19. Wells M. A. A kinetic study of the phospholipase A 2 (Crotalus adamanteus) catalyzed hydrolysis of 1,2-dibutyryl-sn-glycero-3-phosphorylcholine. Biochemistry. 1972 Mar 14;11(6):1030–1041. doi: 10.1021/bi00756a013. [DOI] [PubMed] [Google Scholar]
  20. Williams R. E., Wisnieski B. J., Rittenhouse H. G., Fox C. F. Utilization of fatty acid supplements by cultured animal cells. Biochemistry. 1974 Apr 23;13(9):1969–1977. doi: 10.1021/bi00706a029. [DOI] [PubMed] [Google Scholar]
  21. Wisnieski B. J., Williams R. E., Fox C. F. Manipulation of fatty acid composition in animal cells grown in culture. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3669–3673. doi: 10.1073/pnas.70.12.3669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yang S. F., Freer S., Benson A. A. Transphosphatidylation by phospholipase D. J Biol Chem. 1967 Feb 10;242(3):477–484. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES